BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Hai-Yan Jia,Hong-Li Zhao,Ting Wang,Pin-Ru Chen,Bin-Cheng Yin,Bang-Ce Ye
MicroRNAs (miRNAs) play an essential role in cancer diagnosis and prognosis. Developing a new method for sensitive detection of miRNA is constantly in demand. CRISPR/Cas12a system can nonspecifically cleave single-stranded DNA after specific recognition of target DNA, showing tremendous potential in molecular diagnostics. However, CRISPR-based detection methods require synthesizing different crRNAs for detecting different targets, which limit their widespread application. Herein, we design a versatile and sensitive miRNA detection platform based on CRISPR/Cas12a system combined with a hybridization chain reaction (HCR) circuit. In this design, the HCR circuit as the signal transducer converts each miRNA into multiple DNA duplexes, which act as the activators to activate the trans -cleavage activity of Cas12a for further signal amplification . More importantly, this platform can sensitively detect different miRNAs without changing the spacer sequence of crRNA due to the fixed activators formed by HCR. In addition, the consistency between the proposed platform and RT-qPCR in miRNA detection extracted from different cell lines validated its practicability, demonstrating the potential in clinical diagnosis of cancers and monitoring therapy.
研究领域:
分子生物学
·
生物技术
·
癌症研究
·
诊断技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Bio Materials
·
2019
·
IF 0
Feiyi Wang,Sisi Hu,Qi Sun,Qiang Fei,Chao Ma,Cuifen Lu,Junqi Nie,Zuxing Chen,Jun Ren,Guo-Rong Chen,Guichun Yang,Xiao-Peng He,Tony D James
Currently, chemotherapy is a widely used and important treatment for cancer. However, almost all of the treatments have shortcomings associated with poor specificity and high toxicity, which results in severe side effects to normal cells and tissue. This is a very important problem, and yet, it currently remains unanswered. Therefore, the development of the method for the more effective delivery of anticancer drugs to their targets and real-time monitoring of the localization of the drugs are very important. Herein, we designed a theranostic prodrug: CPT-p-Leu, which was constructed using fluorescent camptothecin (CPT), a self-immolative linker and leucine (Leu) residue. Upon exposure to LAP (leucine aminopeptidase: LAP), the amide bond in CPT-p-Leu will be cleaved, followed by an intramolecular 1,6-elimination, which triggers the active anticancer drug (CPT) release and recovers the fluorescence of CPT. With our design, the anticancer drug, CPT, can be used as both a drug and a fluorescence reporter, making our system suitable to accurately and effectively track the released CPT distribution. Based on this strategy, CPT-p-Leu could achieve the chemoselective detection of LAP and monitoring of the anticancer drug release. Furthermore, it also provides a very convenient way to accurately determine the location of the released drug in living samples. In addition, CPT-p-Leu shows a good cell membrane permeability and enhanced cytotoxicity toward LAP overexpressing cancer cells. We anticipate that our research will facilitate the development of improved theranostic systems for cancer therapy.
研究领域:
肿瘤学
·
药物化学
·
分子生物学
·
药物递送系统
✓ 已验证
产品关联
DOI
PubMed
Journal of Analytical Science and Technology
·
2024
·Vol. 15
·期 1
·1-8
·
IF 2.5
Li Yunxiao,Wang Qiuxia,Wang Yali
The abnormally expressed microRNAs (miRNAs) serve as crucial indicators for disease diagnosis and are strongly associated with the progression of various diseases. The quantification of miRNAs is highly significant for the clinical diagnosis and treatment of various types of cancers. This study utilizes the CRISPR/Cas13a system, which combines CRISPR RNA (crRNA) and CRISPR-associated Cas13a, to recognize the miRNA directly and specifically, thus activating the trans-cleavage activity of Cas13a. By integrating the CRISPR-Cas13a system with an exonuclease-iii (Exo-iii)-assisted chain cleavage of silver ions (Ag+)-aptamer and an Ag+ -based color reaction, a method for the detection of miRNA that is specific and sensitive is developed. This approach demonstrated exceptional sensitivity in detecting miRNA, with a low detection limit of 5.12 fM. Moreover, the suggested method was effectively utilized to precisely measure the amount of miRNA in intricate biological samples, demonstrating its practical applicability. Furthermore, due to its elevated sensitivity and simple probe design and fluorophore labeling, the suggested colorimetric technique holds great potential for use in clinical diagnostics.
研究领域:
分子生物学
·
医学诊断
·
生物技术
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Songkuan Zhuang,Xiaojun Huang,Muyan Diao,Ruiling Dong,Yueyu Dong,Jun Liu,Yizhen Liu
Current one-pot CRISPR diagnostics necessitate meticulous control of nucleic acid hybridization parameters or extensive modification of CRISPR components to achieve complete enzymatic silencing, a fundamental bottleneck limiting their robustness and generalizability. Here, we resolve this challenge through dynamic crRNA reconfiguration, a paradigm-shifting strategy that exploits the intrinsic structural duality of CRISPR RNA. We present CONVERT (CRISPR One-Pot Nucleic acid detection Via Engineered crRNA Reconfiguration Technology), a universal platform where nontarget intact crRNA acts as a universal suppressor, achieving complete Cas12a inhibition during RPA (recombinase polymerase amplification) by irreversible enzyme sequestration. Target detection is initiated through programmable conversion to split crRNA activators, bypassing sequence-specific design constraints entirely. This crRNA conformational switching mechanism is implemented via photocleavable linker cleavage and subsequent assembly with presupplied truncated crRNA (tcrRNA), activating trans-cleavage signal amplification. Three transformative advances emerge: First, the endogenous crRNA engineering eliminates dependency on exogenous nucleic acid blockers or chemical modifications, reducing optimization costs and time. Second, near-total enzymatic suppression ensures zero cross-talk between amplification and detection phases. Third, the unified workflow enables contamination-resistant operation through spatiotemporal control, requiring no physical partitioning. Validated for Enterocytozoon bieneusi detection, the system demonstrates attomolar sensitivity of 1 aM, 100% diagnostic concordance with qPCR, and absolute specificity against related pathogens. By effectively decoupling CRISPR regulation from sequence-context constraints through rational crRNA structural plasticity, this work establishes a generalized framework for next-generation molecular diagnostics.
研究领域:
分子生物学
·
生物工程
·
诊断技术
·
合成生物学
·
核酸扩增
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biomaterials Science
·
2023
·
IF 6.6
Siyi Li,Qinglin Wang,Yingying Ren,Pengfei Zhong,Pengtao Bao,Shanyue Guan,Xiaochen Qiu,Xiaozhong Qu
The development of cancer treatment is of great importance especially in the early stage. In this work, we synthesized pH-sensitive amphiphilic ruthenium complex containing two alkyl chains and two PEG chains which was utilized as oxygen sensitive fluorescent probe for co-assembly with lipids to harvest a liposomal delivery system (RuPC) for the encapsulation of a photothermal agent indocyanine green (ICG). The resultant ICG encapsulated liposome (RuPC@ICG) enabled to deliver ICG into cells via membrane fusion pathway by which the ruthenium complex was localized in the cell membrane for better detecting the extracellular oxygen concentration. Such characteristic endowed the ratiometric imaging feature to distinguish tumour location from normal tissues at just 3 days after cancer cells were implanted by monitoring the hypoxia condition and tracing the metabolism. Moreover, the pH sensitivity of the liposomes favoured the cell uptake, and improved the anti-tumour efficiency of the formulation in vivo under NIR irradiation. Should liposomal systems have less safety issue, our work not only provided a facile way for the construction of theragnostics by combining phototherapy with photoluminescent imaging, but hopefully paving the way for the clinical translation from bench to bedside.
研究领域:
癌症研究
·
纳米医学
·
生物医学工程
·
药学
·
治疗诊断学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BMC CANCER
·
2018
·Vol. 18
·期 1
·1-11
·
IF 3.288
Liu Hai-Ning,Wu Hao,Tseng Yu-Jen,Chen Yan-Jie,Zhang Dan-Ying,Zhu Lin,Dong Ling,Shen Xi-Zhong,Liu Tao-Tao
Background Many novel diagnostic biomarkers have been developed for gastric cancer (GC) recently. We chose two methods with high diagnostic value, the detection of serum microRNAs and metabolomics based on gas chromatography/mass spectrometry (GC/MS), and aimed to establish appropriate models.Methods We reviewed the diagnostic accuracies of all microRNAs identified by previous diagnostic tests. Then appropriate microRNAs and their combinations were validated the diagnostic value. We included 80 patients with GC and 82 healthy controls (HCs) and detected the expression of the microRNAs. GC/MS analysis was conducted, and we used three multivariate statistical analyses to establish diagnostic models. The concentrations of carcinoembryonic antigen (CEA) and carbohydrate antigen 19–9 (CA19–9) were detected for comparison with the novel models.Results Sixty-seven published studies and 70 microRNAs were finally included in the systematic review. MiR-18a, miR-19a, miR-21, miR-92a, miR-199a and miR-421 were chosen to further validate their diagnostic efficiencies. Five of those microRNAs in GC patients had significantly different expression. The combination of miR-19a and miR-92a had the highest area under the curve (AUC) at 0.850 with a sensitivity of 91.3% and a specificity of 61.0%. The GC/MS analysis performed an excellent diagnostic value and the AUC reached 1.0.Conclusion There is a good potential for microRNAs and GC/MS analysis as new diagnostic methods in view of their high diagnostic value compared with traditional biomarkers.
研究领域:
肿瘤学
·
分子生物学
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF THE ENERGY INSTITUTE
·
2022
·
IF 6.47
Xuan Zhao,Dandan Qi,Kaixuan Yang,Chen Chen,Lei Xu,Yaoyao Ying,Tianjiao Li,Dong Liu
This work focused on effects of n- butanol addition on sooting transition process in iso -octane counterflow diffusion flames using optical and chemical diagnostics. Three iso -octane flames were studied with pure iso -octane, a 20% n- butanol addition mixture and a 40% n- butanol addition mixture as fuels. Optical diagnostics was carried out to determine effects of n -butanol addition on the sooting transition process using a high-resolution digital camera based on image processing and transition point extraction methods. Online gas chromatograph was used to investigate the flame chemistry in the sooting transition process. The results showed that the sooting transition point was delayed with the increase of n -butanol addition, which indicated that the addition of n -butanol could suppress the soot formation. To reveal the mechanism for effects of n -butanol on sooting transition process, kinetic simulations were further performed. The experimental results of flame chemistry were compared with the predictions from two mechanisms for iso -octane and n -butanol mixture. Both mechanisms were found to be capable of well capturing the main variations of most species in flames. After n -butanol addition, the changes of fuel-specific primary decomposition products were more obvious. It was noted that the decreased concentration of carbon which was available for benzene formation was the main reason for the delay of sooting transition point with n -butanol additions. Moreover, the inhibition of PAHs growth from n -butanol addition also could contribute to this kind of delayed effects.
研究领域:
燃烧科学
·
化工工程
·
光学诊断
·
火焰化学
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Lifeng Zhang,Shihua Luo,Rui Fan,Ruixi Li,Wenbin Li,Siting Chen,Fei Lan,Yitong Zhu,Tingting Ji,Ye Zhang,Ling Li
APE1, an essential enzyme for DNA repair, is overexpressed in various cancers and has been identified as a potential biomarker for cancer diagnosis. However, detecting APE1 at low expression levels in the early stage of cancer presents a significant obstacle. Here, we introduced a novel localized Cas12a-based cascade amplification (LCas12a-CA) method. This method confined both the terminal deoxynucleotidyl transferase and the crRNA/Cas12a complex onto the surfaces of gold nanoparticles (AuNPs). This confinement not only boosts the stability of the multiple enzymes but also induces a substrate channeling effect. As a result, it significantly accelerates the reaction rate and enhances the sensitivity of APE1 detection. Upon the addition of APE1, the AP sites within the APE1 primer can be recognized and cleaved by APE1, exposing the 3′-OH ends. In the presence of LCas12a-CA, polyA sequences are generated at 3′-OH ends with the help of TdT and dATP. The sequences directly enter the Cas12a system, activating the trans -cleavage activity of Cas12a, thereby cutting the reporters on the surface of AuNPs and releasing fluorescence. Our platform demonstrates a detection limit (LOD) as low as 2.51 × 10 −6 U/mL, which is more than 60 times lower than that of free Cas12a-CA. Furthermore, the LCas12a-CA exhibits enhanced resistance ability in extreme environments and has been proven effective for the detection of APE1 in clinical samples. Overall, this work offers a promising platform for robust biosensing in cancer diagnosis and prognosis.
研究领域:
分子生物学
·
癌症研究
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Machine Intelligence
·
2021
·Vol. 3
·期 3
·199-217
·
IF 15.508
Roberts Michael,Driggs Derek,Thorpe Matthew,Gilbey Julian,Yeung Michael,Ursprung Stephan,Aviles-Rivero Angelica I.,Etmann Christian,McCague Cathal,Beer Lucian,Weir-McCall Jonathan R.,Teng Zhongzhao,Gkrania-Klotsas Effrossyni,Rudd James H. F.,Sala Evis,Schönlieb Carola-Bibiane
Machine learning methods offer great promise for fast and accurate detection and prognostication of coronavirus disease 2019 (COVID-19) from standard-of-care chest radiographs (CXR) and chest computed tomography (CT) images. Many articles have been published in 2020 describing new machine learning-based models for both of these tasks, but it is unclear which are of potential clinical utility. In this systematic review, we consider all published papers and preprints, for the period from 1 January 2020 to 3 October 2020, which describe new machine learning models for the diagnosis or prognosis of COVID-19 from CXR or CT images. All manuscripts uploaded to bioRxiv, medRxiv and arXiv along with all entries in EMBASE and MEDLINE in this timeframe are considered. Our search identified 2,212 studies, of which 415 were included after initial screening and, after quality screening, 62 studies were included in this systematic review. Our review finds that none of the models identified are of potential clinical use due to methodological flaws and/or underlying biases. This is a major weakness, given the urgency with which validated COVID-19 models are needed. To address this, we give many recommendations which, if followed, will solve these issues and lead to higher-quality model development and well-documented manuscripts.
研究领域:
医学影像学
·
人工智能
·
流行病学
✓ 已验证
DOI
高影响因子
TALANTA
·
2021
·
IF 6.057
Jing Zhang,Xinxin Wen,Yuting Li,Jing Zhang,Xian Li,Cheng Qian,Yonghui Tian,Rui Ling,Yixiang Duan
Thyroid cancer is a malignant disease with dramatically low advanced-stage 10-year survival. Meanwhile, the metabolites in saliva are becoming a wealthy source of disease biomarkers. However, there is a lack of non-invasive analytical methods for the identification of biomarkers in saliva for the preoperative diagnosis of thyroid cancer. Therefore, we developed an ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) method to simultaneously determine the metabolic levels of 10 amino acids in saliva, aiming to study the amino acid metabolism profile to promote early diagnosis of thyroid cancer. We tested unstimulated whole saliva from patients with papillary thyroid carcinoma (PTC; n = 61) and healthy controls (HC; n = 61), and used receiver operating characteristic (ROC) curves to establish the diagnostic value of potential markers. The method validation results showed good precision, linearity (R 2 > 0.99), recovery (92.2 %–110.3 %), intra- and inter-day precision (RSD < 7 % and RSD < 9 %, respectively). The concentration of 10 amino acids was significantly different between PTC and HC in human salivary analysis (P < 0.05), the area under the curve (AUC) values of a single marker for the diagnosis of PTC were ranging from 0.678 to 0.833. A panel of alanine , valine , proline , phenylalanine was selected in combination yielded the AUC of 0.936, which will improve the accuracy of early diagnosis of thyroid cancer (sensitivity: 91.2 %; specificity: 85.2 %). This study proved the possibility of salivary amino acid biomarkers for PTC early diagnosis, providing a simple auxiliary way for the non-invasive diagnosis of thyroid cancer.
研究领域:
医学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY
·
2022
·
IF 5.062
Lanlan Lin,Haiyang Li,Zekun Chen,Xinyu Wen,Rui Yan,Lei Liu,Caihong Tao
The combination of chemotherapy and photothermal therapy can provide better tumor treatment effect than monotherapy , while the combination of imaging technology can achieve diagnostic effect during the treatment. In this paper, biocompatible mesoporous polydopamine (MPDA) were combined with the near-infrared fluorescent dye IR780 under alkaline condition to form an integrated drug delivery platform for tumor diagnosis and photothermal co-therapy. The particles size and mesoporous pore size of the nanospheres were adjusted by changing the preparation conditions of MPDA, so the platform had strong permeability and large drug loading capacity in tumor cells as drug carriers. The loading and release results of doxorubicin hydrochloride (DOX) proved that the carrier had a high drug loading and sustained release rate, as well as a faster dependent release under acidic and lighting conditions. Photothermal experiments showed that the carrier had high photothermal conversion efficiency, which was very critical in its application in photothermal therapy and fluorescence imaging of tumors. The vitro cytotoxicity test evaluated the survival rates of LX-2 cells and HepG2 cells after drug loading. The results showed that the MPDA@IR780-DOX + NIR carrier had a significant inhibitory effect on HepG2 cells (12.57%), but had little effect on LX-2 cells, which was beneficial for its application in integration of biological diagnosis and treatment.
研究领域:
生物医学工程
·
纳米医学
·
肿瘤学
✓ 已验证
产品关联
DOI
高影响因子
Discover Oncology
·
2024
·Vol. 15
·期 1
·1-10
·
IF 2.8
Fan Ruizhi,Xu Teng,Kuang Yuting
Background Colorectal cancer, which originates from the human colon or rectum, is one of the leading causes of death worldwide. Timely diagnosis and interventional therapy can significantly improve the prognostic survival of colorectal cancer patients, making regular screening and early detection essential. Aim To investigate the regulatory function of lncRNA CTBP1-DT (CTBP1-DT) on colorectal cancer cells and to assess its diagnostic significance. Methods A total of 102 patients with colorectal cancer and 92 healthy individuals were selected. The levels of CTBP1-DT and microRNA-30a-5p (miR-30a-5p) in serum and cell samples of the above subjects were compared by RT-qPCR. The effects of CTBP1-DT and miR-30a-5p dysregulation on the biological functions of colorectal cancer cells were analyzed via CCK-8, flow cytometry and Transwell assays. In addition, the ability of CTBP1-DT and miR-30a-5p to early identify colorectal cancer patients was determined through ROC curve. Results Serum CTBP1-DT was elevated in patients with colorectal cancer, which was obviously higher than in healthy controls. The expression of serum miR-30a-5p was downregulated in colorectal cancer. Both CTBP1-DT and miR-30a-5p have the value of distinguishing colorectal cancer, and the combined diagnostic ability is higher. Knockdown of CTBP1-DT directly targeted miR-30a-5p to repress cell activity and metastatic ability, whereas deregulation of miR-30a-5p eliminated the above inhibitory effects. Conclusion Overexpression of CTBP1-DT has a certain application potential in the diagnosis of colorectal cancer and may be a therapeutic target for colorectal cancer.
研究领域:
肿瘤学
·
分子生物学
·
癌症生物学
·
诊断
✓ 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）