Scientific Reports
·
2017
·Vol. 7
·期 1
·1-9
·
IF 4.259
Li Lanhui,Jin Mingliang,Sun Chenglong,Wang Xiaoxue,Xie Shuting,Zhou Guofu,van den Berg Albert,Eijkel Jan C. T.,Shui Lingling
DNA fragmentation down to a precise fragment size is important for biomedical applications, disease determination, gene therapy and shotgun sequencing. In this work, a cheap, easy to operate and high efficiency DNA fragmentation method is demonstrated based on hydrodynamic shearing in a bubbling system. We expect that hydrodynamic forces generated during the bubbling process shear the DNA molecules, extending and breaking them at the points where shearing forces are larger than the strength of the phosphate backbone. Factors of applied pressure, bubbling time and temperature have been investigated. Genomic DNA could be fragmented down to controllable 1–10 Kbp fragment lengths with a yield of 75.30–91.60%. We demonstrate that the ends of the genomic DNAs generated from hydrodynamic shearing can be ligated by T4 ligase and the fragmented DNAs can be used as templates for polymerase chain reaction. Therefore, in the bubbling system, DNAs could be hydrodynamically sheared to achieve smaller pieces in dsDNAs available for further processes. It could potentially serve as a DNA sample pretreatment technique in the future.
研究领域:
生物技术
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Luke Wei,Yuxin Hu,Jiamin Wu,Jing Mao,Tianqin Yin,Jieqiong Qiu
Doxorubicin (DOX), a widely utilized chemotherapeutic antibiotic, increasingly threatens environmental and public health due to its persistence in marine ecosystems, bioaccumulation in organisms, and contamination of agricultural systems. In response to the urgent demand for effective DOX detection, we have developed a label-free fluorescent aptamer biosensor (FAB) that employs SYBR Green I (SG I)-mediated signal amplification. A key innovation in this design is the strategic fragmentation of the 41-mer complementary strand (CP) of the DOX aptamer into three 11-mer fragments. It significantly mitigates nonspecific SG I binding interference caused by the longer ssDNA CP, enhancing the fluorescence signal ratio (F dsDNA /F ssDNA ) from 4.6-fold to 12.6-fold. Upon DOX introduction, competitive binding between DOX, SG I, and DNA further suppresses background noise, yielding an exceptional fluorescence quenching ratio (F 0 /F) of 52.8. The optimized FAB demonstrates ultra-sensitive DOX detection, with a detection limit of 1.41 nM and a linear response between 1.41 nM and 300 nM. The recoveries of DOX spiked into pond water and drinking water samples ranged from 95.1% to 101.8%, with relative standard deviations (RSDs) below 3.46%, demonstrating negligible matrix effects and confirming the FAB method’s robustness for environmental monitoring applications. This strategy synergistically enhances signal transduction efficiency, and strengthens detection reliability, all while maintaining operational simplicity. As a result, this FAB for DOX detection holds transformative potential, poised to revolutionize biosensor technology in the fields of food safety, environmental monitoring, and biosecurity.
研究领域:
生物传感器
·
环境科学
·
分析化学
·
分子生物学
·
生物化学
·
纳米技术
·
公共卫生
·
海洋生态学
✓ 已验证
DOI
PubMed
高影响因子
Metallomics
·
2021
·Vol. 13
·期 10
·
IF 4.526
Ding Haoxuan,Zhang Qian,Yu Xiaonan,Chen Lingjun,Wang Zhonghang,Feng Jie
Iron overload is an important contributor to disease. The liver, the major site of iron storage in the body, is a key organ impacted by iron overload. While several studies have reported perturbations in liver lipids in iron overload, it is not clear, on a global scale, how individual liver lipid ions are altered. Here, we used lipidomics to study the changes in hepatic lipid ions in iron-overloaded mice. Iron overload was induced by daily intraperitoneal injections of 100 mg/kg body weight iron dextran for 1 week. Iron overload was verified by serum markers of iron status, liver iron quantitation, and Perls stain. Compared with the control group, the serum of iron-overload mice exhibited low levels of urea nitrogen and high-density lipoprotein (HDL), and high concentrations of total bile acid, low-density lipoprotein (LDL), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH), suggestive of liver injury. Moreover, iron overload disrupted liver morphology, induced reactive oxygen species (ROS) production, reduced superoxide dismutase (SOD) activity, caused lipid peroxidation, and led to DNA fragmentation. Iron overload altered the overall composition of lipid ions in the liver, with significant changes in over 100 unique lipid ions. Notably, iron overload selectively increased the overall abundance of glycerolipids and changed the composition of glycerophospholipids and sphingolipids. This study, one of the first to report iron-overload induced lipid alterations on a global lipidomics scale, provides early insight into lipid ions that may be involved in iron overload-induced pathology.
研究领域:
肝病学
·
脂质组学
·
病理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2024
·
IF 2.7
Shuwei Shen,Binbin Huang,Zhiyi Peng,Luting Yu,Lili Ren,Jia Liu,Guoguang Chen
Traditional Chinese medicine (TCM), as a significant complementary and alternative approach, has received increasing attention from medical practitioners worldwide and is now regarded as an important source for modern drug discovery owing to its special therapeutic efficiency and few side effects. In this work, a popular TCM, Ganoderma lingzhi (G. lingzhi), was used as a test drug to show how Raman spectroscopy combined with traditional chemical and biological assays could be used to monitor alternations in molecular events in G. lingzhi extract-induced cells with high sensitivity. The observations indicated that the Raman fingerprint of MDA-MB-231 cells after G. lingzhi triterpene treatment was significantly different in contrast to that of the control, and changes in the Raman bands showed that some important molecular events could be discerned, including protein denaturation, DNA fragmentation, and lipid metabolism disorder. Network pharmacology analysis further supplemented the drug's target and signaling pathways. Our work proposes an effective way for screening potential new drug products early from the complex chemical compositions of TCM, as well as for accelerating preclinical medicinal chemistry optimization cycles, playing a significant role in the development of new drugs.
研究领域:
药理学
·
光谱学
·
细胞生物学
·
中药
·
药物发现
✓ 已验证
产品关联
DOI
高影响因子
Food Bioscience
·
2019
·
IF 3.22
Chun-Min Ma,Tie-Jing Li,Xin-Huai Zhao
Lactoferrin hydrolysates (LF-H) were digested using pepsin and modified using a pepsin-catalyzed plastein reaction with tryptophan (Trp) to obtain a target product. LF-H, the target product, plastein reaction-modified LF-H alone, and a mixture of LF-H and Trp were evaluated for their activities with human gastric cancer BGC-823 cells at 24 and 48 h. Assay results indicated that the target product had decreased free amino groups and had incorporated Trp. LF-H and the plastein reaction-modified LF-H alone did not show detectable growth inhibition on BGC-823 cells at 1–3 mg/mL; however, the target product at the same dose levels inhibited cell growth, and showed higher activity than the mixture ( P < 0.05). The target product at 2 and 3 mg/mL stopped cell cycle progression at the G0/G1-phase, induced DNA fragmentation , and showed apoptosis induction, which increased the number of apoptotic cells. Moreover, time- and dose-dependent responses were observed in these experiments. RT-PCR assay results indicated that exposure of the cells to the target product resulted in up- and down-regulated mRNA expression for 7 apoptosis-related genes, namely Bcl-2, Bcl-xl, Bad, Bax, caspase-3, caspase-6, and caspase-9. Thus, the plastein reaction increased in vitr o anticancer activities of LF-H with BGC-823 cells, and more importantly, both the plastein reaction and Trp incorporation contributed to enhanced activities.
研究领域:
生物化学
·
癌症研究
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
Molecular Medicine Reports
·
2015
·Vol. 12
·期 3
·3869-3873
·
IF 1.554
Shi‑Min Yu,Dong‑Hui Hu,Jian‑Jun Zhang
Hepatocellular carcinoma (HCC) is a highly malignant tumor, associated with poor patient prognoses, and high rates of morbidity and mortality. To date, the therapeutic strategies available for the treatment of HCC remain limited. The present study aimed to elucidate the anticancer activity of umbelliferone, a naturally occurring coumarin derivative isolated from Ferula communis, against the HepG2 HCC cell line. A 3‑(4,5‑dimthylthaizol‑2‑yl)‑2,5, diphenyltetrazolium bromide assay was used to evaluate cell viability following umbelliferone treatment, and the effects of umbelliferone on cell cycle progression and apoptosis were evaluated using flow cytometry. The presence of morphological features characteristic of apoptosis, including cell shrinkage, membrane blebbing, nuclear condensation and apoptotic body formation, were evaluated in HepG2 cells following umbelliferone treatment. Cell cycle analysis conducted via propidium iodide (PI) staining indicated that umbelliferone treatment induced cell cycle arrest at S phase in HepG2 cells. Analysis with Annexin V and PI staining revealed that umbelliferone induced apoptotic events in HepG2 cells in a concentration‑dependant manner (0‑50 µM). Umbelliferone also induced dose‑dependant DNA fragmentation. In conclusion, umbelliferone was found to exhibit significant anticancer effects via the induction of apoptosis, cell cycle arrest and DNA fragmentation in HepG2 cancer cells.
研究领域:
肿瘤学
·
药理学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANIMAL REPRODUCTION SCIENCE
·
2025
·
IF 3.3
Chaoyue Cao,Xue Qin,Yiwen Sha,Haoqi Xiao,Zhenheng Lai,Yi Zheng,Rui Cai,Weijun Pang
Vitamin E (VE) is widely used as an antioxidant in boar semen preservation. Its low water solubility and limited bioavailability, however, compromise its functional efficacy during storage. To address these limitations, we developed a lecithin-stabilized vitamin E nanoemulsion (NE) to enhance its dispersibility and antioxidant efficacy. Semen from Duroc boars was diluted in Beltsville thawing solution (BTS) containing either 0.6 mM NE, 4 mM free VE, or no additive, and stored at 17 °C. Compared to VE and control groups, NE significantly enhanced sperm motility, preserved the structural integrity of the plasma and acrosomal membranes, and maintained mitochondrial function, while reducing oxidative stress and DNA fragmentation over a 7d storage period ( P < 0.01). Untargeted metabolomics revealed that NE supplementation regulated key pathways including unsaturated fatty acid metabolism and glutathione-related antioxidation. Artificial insemination (AI) trials involving 637 sows confirmed that NE-treated semen improved total and healthy piglet numbers per litter ( P < 0.05), without adverse effects. These findings suggest that NE provides an effective and practical strategy for enhancing boar semen preservation and reproductive outcomes in porcine production.
研究领域:
动物繁殖
·
纳米技术
·
抗氧化研究
·
代谢组学
·
兽医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
WATER RESEARCH
·
2020
·
IF 9.13
Hangbo Xu,Yupan Zhu,Mengru Du,Yuqi Wang,Siyao Ju,Ruonan Ma,Zhen Jiao
Although the identification of effective reactive oxygen species (ROS) generated by plasma has been extensively studied, yet the subcellular mechanism of microbial inactivation has never been clearly elucidated in plasma disinfection processes. In this study, subcellular mechanism of yeast cell inactivation during plasma-liquid interaction was revealed in terms of comprehensive factors including cell morphology, membrane permeability, lipid peroxidation, membrane potential, intracellular redox homeostasis (intracellular ROS and H 2 O 2 , and antioxidant system (SOD, CAT and GSH)), intracellular ionic equilibrium (intracellular H + and K + ) and energy metabolism (mitochondrial membrane potential, intracellular Ca 2+ and ATP level). The ROS analysis show that ·OH, 1 O 2 , ·O 2 − and H 2 O 2 were generated in this plasma-liquid interaction system and ·O 2 − served as the precursor of 1 O 2 . Additionally, the solution pH was reduced. Plasma can effectively inactivate yeast cells mainly via apoptosis by damaging cell membrane, intracellular redox and ion homeostasis and energy metabolism as well as causing DNA fragmentation. ROS scavengers ( l -His, d -Man and SOD) and pH buffer (phosphate buffer solution, PBS) were employed to investigate the role of five antimicrobial factors (·OH, 1 O 2 , ·O 2 − , H 2 O 2 and low pH) in plasma sterilization. Results show that they have different influences on the aforementioned cell physiological activities. The ·OH and 1 O 2 contributed most to the yeast inactivation. The ·OH mainly attacked cell membrane and increased cell membrane permeability. The disturb of cell energy metabolism was mainly attributed to 1 O 2 . The damage of cell membrane as well as extracellular low pH could break the intracellular ionic equilibrium and further reduce cell membrane potential. The remarkable increase of intracellular H 2 O 2 was mainly due to the influx of extracellular H 2 O 2 via destroyed cell membrane, which played a little role in yeast inactivation during 10-min plasma treatment. These findings provide comprehensive insights into the antimicrobial mechanism of plasma, which can promote the development of plasma as an alternative water disinfection strategy.
研究领域:
微生物学
·
等离子体物理
·
环境科学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2019
·
IF 3.571
Jinming Wang,Jiarong Yang,Xiaofang Cheng,Rui Xiao,Yangfei Zhao,Huimiao Xu,Yaya Zhu,Zipeng Yan,Mohammad Mehdi Ommati,Ram Kumar Manthari,Jundong Wang
Excessive fluoride mainly causes skeletal lesions. Recently, it has been reported that an appropriate level of calcium can alleviate fluorosis. However, the appropriate concentration and mechanism of calcium addition is unclear. Hence, we evaluated the histopathology and ultrastructure, DNA fragmentation, hormonal imbalances, biomechanical levels, and expression of apoptosis-related genes after treating the rats with 150 mg/L NaF and different concentrations of CaCO3. Our results suggested that NaF induced the histopathological and ultrastructural injury, with a concomitant increase in the DNA fragmentation (P < 0.05) and serum OC (17.5 ± 0.89 pmoL/L) at 120 days. In addition, the qRT-PCR and western blotting results indicated that NaF exposure upregulated the mRNA and protein expression of Bax, Calpain, Caspase 12, Caspase 9, Caspase 7, Caspase 3, CAD, PARP, and AIF while downregulated Bcl-2 (P < 0.01) and decreased the bone ultimate load by 27.1%, the ultimate stress by 10.1%, and the ultimate deformity by 23.3% at 120 days. However, 1% CaCO3 supplementation decreased the serum OC (14.7 ± 0.65 pmoL/L), bone F content (P < 0.01), and fracture and breakage of collagen fibers and changed the expression of endoplasmic reticulum pathway-related genes and proteins at 120 days. Further, 1% CaCO3 supplementation increased the bone ultimate load by 20.9%, the ultimate stress by 4.89%, and the ultimate deformity by 21.6%. In summary, we conclude that 1% CaCO3 supplementation alleviated fluoride-induced bone damage by inhibiting endoplasmic reticulum stress and mitochondrial dysfunction.
研究领域:
药理学
·
毒理学
·
生物学
·
病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2022
·
IF 8.786
Xie Yiwei,Liang Hongrui,Jiang Ning,Liu Dingyuan,Zhang Naiwen,Li Qilong,Zhang Kai,Sang Xiaoyu,Feng Ying,Chen Ran,Zhang Yiwei,Chen Qijun
Trypanosoma brucei, the pathogen causing African sleeping sickness (trypanosomiasis) in humans, causes debilitating diseases in many regions of the world, but mainly in African countries with tropical and subtropical climates. Enormous efforts have been devoted to controlling trypanosomiasis, including expanding vector control programs, searching for novel anti-trypanosomial agents, and developing vaccines, but with limited success. In this study, we systematically investigated the effect of graphene quantum dots (GQDs) on trypanosomal parasites and their underlying mechanisms. Ultrasmall-sized GQDs can be efficiently endocytosed by T. brucei and with no toxicity to mammalian-derived cells, triggering a cascade of apoptotic reactions, including mitochondrial disorder, intracellular reactive oxygen species (ROS) elevation, Ca2+ accumulation, DNA fragmentation, adenosine triphosphate (ATP) synthesis impairment, and cell cycle arrest. All of these were caused by the direct interaction between GQDs and the proteins associated with cell apoptosis and anti-oxidation responses, such as trypanothione reductase (TryR), a key protein in anti-oxidation. GQDs specifically inhibited the enzymatic activity of TryR, leading to a reduction in the antioxidant capacity and, ultimately, parasite apoptotic death. These data, for the first time, provide a basis for the exploration of GQDs in the development of anti-trypanosomials.
研究领域:
寄生虫学
·
纳米技术
·
分子生物学
·
药理学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF FOOD SCIENCE
·
2022
·Vol. 87
·期 9
·4119-4136
·
IF 3.693
Huxuan Wang,Zhonghua Peng,Cong Wang,Yanan Zhu,Fei Xia,Hongmin Sun
This work aimed for the first time to provide detailed insights into thymol and trans -cinnamaldehyde's mechanisms of action on the food-spoilage yeast Zygosaccharomyces rouxii and offers evidence in favor of the activation of an apoptosis-like phenotype. The action mechanisms of thymol and trans -cinnamaldehyde were investigated by the measurement of a series of typical apoptotic features using flow cytometer or microplate reader. Moreover, quantitative reverse transcription PCR (QRT-PCR) was performed to investigate the effects of thymol and trans -cinnamaldehyde on the transcription of key regulators of apoptosis in Z. rouxii . The results indicated that the treatment of Z. rouxii with thymol or trans -cinnamaldehyde (minimum inhibitory and subinhibitory concentrations) triggered reactive oxygen species (ROS) accumulation, elevated intracellular Ca 2+ level, and depolarized mitochondrial membrane potential (MMP) coupled with hallmarks of apoptosis including mitochondrial cytochrome c (cyt c) release, metacaspase activation, phosphatidylserine (PS) exposure, and DNA fragmentation. Moreover, thymol or trans -cinnamaldehyde treatment upregulated the transcription of proapoptotic regulators including Yca 1, Dnm 1, Nuc 1, Ndi 1, and Mmi 1 and downregulated the transcription of antiapoptotic regulators of Fis 1 and Cdc 48, further confirming the apoptosis induced by thymol and trans -cinnamaldehyde in Z. rouxii . In summary, thymol and trans -cinnamaldehyde probably induced apoptosis through a metacaspase-dependent mitochondrial pathway in Z. roxuii . These findings provide theoretical support for the development of safe and efficient agents used in food preservation. Practical Application The results will provide a new idea for the systematic analysis of the antifungal mechanisms of thymol and trans -cinnamaldehyde and also provide a theoretical support for the development and application of natural food preservatives, which is of positive significance for the effective control of the spoilage caused by Z. rouxii in food processing and storage and the protection of consumers’ health.
研究领域:
食品科学
·
微生物学
·
药理学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ENVIRONMENTAL SCIENCE & TECHNOLOGY
·
2018
·
IF 6.653
Chang Wang,Xue Yang,Qi Zheng,Birget Moe,Xing-Fang Li
The developmental toxicity of water disinfection byproducts remains unclear. Here we report the study of halobenzoquinone (HBQ)-induced in vivo developmental toxicity and oxidative stress using zebrafish embryos as a model. Embryos were exposed to 0.5–10 μM of individual HBQs and 0.5–5 mM haloacetic acids for up to 120 h postfertilization (hpf). LC50 values of the HBQs at 24 hpf were 4.6–9.8 μM, while those of three haloacetic acids were up to 200 times higher at 1900–2600 μM. HBQ exposure resulted in significant developmental malformations in larvae, including failed inflation of the gas bladder, heart malformations, and curved spines. An increase in reactive oxygen species was observed, together with a decrease in superoxide dismutase activity and glutathione content. Additionally, the antioxidant N-acetyl-l-cysteine significantly mitigated all HBQ-induced effects, supporting that oxidative stress contributes to HBQ toxicity. Further experiments examined HBQ-induced effects on DNA and genes. HBQ exposure increased 8-hydroxydeoxyguanosine levels, DNA fragmentation, and apoptosis in larvae, with apoptosis induction related to changes in the gene expression of p53 and mdm2. These results suggest that HBQs are acutely toxic, causing oxidative damage and developmental toxicity to zebrafish larvae.
研究领域:
毒理学
·
环境科学
·
发育生物学
·
分子生物学
✓ 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）