Co-Delivery of Gemcitabine and Honokiol by Lipid Bilayer-Coated Mesoporous Silica Nanoparticles Enhances Pancreatic Cancer Therapy via Targeting Depletion of Tumor Stroma.

Syndecan-1 (SDC1) modified lipid bilayer (LB)-coated mesoporous silica nanoparticles (MSN) to co-deliver gemcitabine (GEM) and honokiol (HNK) were prepared for the targeting treatment of pancreatic cancer. The encapsulation efficiencies of GEM and HNK in SDC1-LB-MSN-GEM/HNK were determined to be 60.3 ± 3.2% and 73.0 ± 1.1%. The targeting efficiency of SDC1-LB-MSN-GEM/HNK was investigated in BxPC-3 cells in vitro. The fluorescence intensity in the cells treated with SDC1-LB-MSN-Cou6 was 2-fold of LB-MSN-Cou6-treated cells, which was caused by SDC1/IGF1R-mediated endocytosis. As anticipated, its cytotoxicity was significantly increased. Furthermore, the mechanism was verified that SDC1-LB-MSN-HNK induced tumor cell apoptosis through the mitochondrial apoptosis pathway. Finally, the biodistribution, tumor growth inhibition, and preliminary safety studies were performed on BALB/c nude mice bearing BxPC-3 tumor models. The tumor growth inhibition index of SDC1-LB-MSN-GEM/HNK was 56.19%, which was 1.45-fold and 1.33-fold higher than that of the free GEM/HNK and LB-MSN-GEM/HNK treatment groups, respectively. As a result, SDC1-LB-MSN-GEM/HNK combined advantages of both GEM and HNK and simultaneously targeted and eliminated pancreatic cancerous and cancer-associated stromal cells. In summary, the present study demonstrated a new strategy of synergistic GEM and HNK to enhance the therapeutic effect of pancreatic cancer via the targeting depletion of tumor stroma.

Polysialic acid-functionalized liposomes for efficient honokiol delivery to inhibit breast cancer growth and metastasis.

To improve the anti-metastasis effects of honokiol (HNK) on breast cancer, we designed cationic liposomes (Lip) in which HNK was encapsulated into Lip, and its surface was modified with negatively charged polysialic acid (PSA-Lip-HNK) for efficient treatment of breast cancer. PSA-Lip-HNK possessed a homogeneous spherical shape and high encapsulation efficiency. In vitro 4T1 cell experiments indicated that PSA-Lip-HNK increased cellular uptake and cytotoxicity via the endocytosis pathway mediated by PSA and selectin receptors. Furthermore, the significant antitumor metastasis impact of PSA-Lip-HNK was confirmed by wound healing and cell migration and invasion. Enhanced in vivo tumor accumulation of the PSA-Lip-HNK was observed in 4T1 tumor-bearing mice by living fluorescence imaging. For in vivo antitumor experiments using 4T1 tumor-bearing mice, PSA-Lip-HNK exhibited a higher tumor growth and metastasis inhibition compared with unmodified liposomes. Therefore, we believe that PSA-Lip-HNK well combined biocompatible PSA nano-delivery and chemotherapy, providing a promising drug delivery approach for metastatic breast cancer therapy.

Metagenomic next-generation sequencing to identify pathogens and cancer in lung biopsy tissue.

BACKGROUND: Lung biopsy tissue samples can be used for infection detection and cancer diagnosis. Metagenomic next-generation sequencing (mNGS) has the potential to further improve diagnosis. METHODS: From July 2018 to May 2020, lung biopsy samples of 133 patients with suspected pulmonary infection or abnormal imaging findings were collected and subjected to clinical microbiological testing, Illumina and Nanopore sequencing to identify pathogens. The neural networks were pretrained by extracting features of human reads from 2,095 metagenomic next-generation sequencing results, and the human reads of lung biopsy samples were entered into the validated pipeline to predict the risk of cancer. FINDINGS: Based on the pathogen-cancer detection pipeline, the Illumina platform showed 77·6% sensitivity and 97·6% specificity compared to the composite reference standard for infection diagnosis. However, the Nanopore platform showed 34·7% sensitivity and 98·7% specificity. mNGS identified more fungi, which was confirmed by subsequent pathological examination. M. tuberculosis complex was weakly detected. For cancer detection, compared with histology, the Illumina platform showed 83·7% sensitivity and 97·6% specificity, diagnosing an additional 36 cancer patients, of whom half had abnormal imaging findings (pulmonary shadow, space-occupying lesions, or nodules). INTERPRETATION: For the first time, we have established a pipeline to simultaneously detect pathogens and cancer based on Illumina sequencing of lung biopsy tissue. This pipeline efficiently diagnosed cancer in patients with abnormal imaging findings. FUNDING: This work was supported by the National Key Research and Development Program of China and National Natural Science Foundation of China.

Clinical Profile, Prognostic Factors, and Outcome Prediction in Hospitalized Patients With Bloodstream Infection: Results From a 10-Year Prospective Multicenter Study.

Background: Bloodstream infection (BSI) is one of the most common serious bacterial infections worldwide and also a major contributor to in-hospital mortality. Determining the predictors of mortality is crucial for prevention and improving clinical prognosis in patients with nosocomial BSI. Methods: A nationwide prospective cohort study was conducted from 2007 until 2016 in 16 teaching hospitals across China. Microbiological results, clinical information, and patient outcomes were collected to investigate the pathogenic spectrum and mortality rate in patients with BSI and identify outcome predictors using multivariate regression, prediction model, and Kaplan-Meier analysis. Results: No significant change was observed in the causative pathogen distribution during the 10-year period and the overall in-hospital mortality was 12.83% (480/3,741). An increased trend was found in the mortality of patients infected with Pseudomonas aeruginosa or Acinetobacter baumannii, while a decreased mortality rate was noted in Staphylococcus aureus-related BSI. In multivariable-adjusted models, higher mortality rate was significantly associated with older age, cancer, sepsis diagnosis, ICU admission, and prolonged hospital stay prior to BSI onset, which were also determined using machine learning-based predictive model achieved by random forest algorithm with a satisfactory performance in outcome prediction. Conclusions: Our study described the clinical and microbiological characteristics and mortality predictive factors in patients with BSI. These informative predictors would inform clinical practice to adopt effective therapeutic strategies to improve patient outcomes.

Clinical and microbiological characteristics of adults with hospital-acquired pneumonia: a 10-year prospective observational study in China.

Hospital-acquired pneumonia (HAP) is a significant nosocomial infection; data on the distribution and antimicrobial resistance profiles of HAP in China are limited. We included 2827 adult patients with HAP from the Chinese Antimicrobial Resistance Surveillance of Nosocomial Infections network admitted in 15 Chinese teaching hospitals between 2007 and 2016. Clinical data and antimicrobial susceptibility of isolated pathogens were obtained from the medical records and central laboratory, respectively. Multivariable logistic regression was performed to determine the risk factors for mortality and multidrug resistance (MDR). A total of 386 (13.7%) patients died in the hospital, while 1181 (41.8%) developed ventilator-associated pneumonia (VAP). Active immunosuppressant therapy (OR 1.915 (95% CI 1.475-2.487)), solid tumor (OR 1.860 (95% CI 1.410-2.452)), coma (OR 1.783 (95% CI 1.364-2.333)), clinical pulmonary infection score ≥7 (OR 1.743 (95% CI 1.373-2.212)), intensive care unit stay (OR 1.652 (95% CI 1.292-2.111)), age ≥65 years (OR 1.621 (95% CI 1.282-2.049)), and tracheal cannula insertion (OR 1.613 (95% CI 1.169-2.224)) were independent risk factors for in-hospital mortality. Liver cirrhosis (OR 3.120 (95% CI 1.436-6.780)) and six other variables were independent predictors of MDR. Acinetobacter baumannii (25.6%), Pseudomonas aeruginosa (20.1%), Klebsiella pneumoniae (15.4%), and Staphylococcus aureus (12.6%) were the most common pathogens (MDR prevalence 64.9%). Isolates from VAP patients showed more A. baumannii and less K. pneumoniae and E. coli strains (p < 0.001, respectively) than those from patients without VAP. The proportion of methicillin-resistant S. aureus strains decreased; that of carbapenem-resistant A. baumannii and Enterobacterales strains increased. There had been changes in the antibiotic resistance profiles of HAP pathogens in China. Risk factors for mortality and MDR are important for the selection of antimicrobials for HAP in China.

Base-enhanced electrochemical water oxidation by a nickel complex in neutral aqueous solution.

A nickel complex, [Ni(TMC)(CH3CN)](NO3)2 (TMC = 1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetradecane, 1), was found to be an efficient water oxidation catalyst in pH 7 phosphate buffer. It exhibits pseudo first order kinetics in electrochemical water oxidation with a catalytic rate of 9.95 s-1, the highest rate for nickel WOCs at neutral pH. Complex 1 also shows superior catalytic activity with respect to that of a copper analogue, [Cu(TMC)(H2O)](NO3)2, under the same conditions. Kinetic studies indicate a more than one order of magnitude rate acceleration with the added bases due to an atom-proton transfer (APT) pathway.

Bifunctional nickel oxide-based nanosheets for highly efficient overall urea splitting.

Porous and hollow Ni0.9Fe0.1Ox microspheres assembled from 2D nanosheets exhibit excellent bifunctional activity for both urea oxidation reaction and hydrogen evolution reaction, in which the potential for overall urea splitting is 1.455 V at 10 mA cm-2.

Detection of Pulmonary Infectious Pathogens From Lung Biopsy Tissues by Metagenomic Next-Generation Sequencing.

Metagenomic next-generation sequencing (mNGS) is a comprehensive approach for sequence-based identification of pathogenic microbes. However, reports on the use of mNGS in pulmonary infection applied to lung biopsy tissues remain scarce. In this study, we applied mNGS to detect the presence of pathogenic microbes in lung biopsy tissues from 20 patients with pulmonary disorders indicating possible infection. We applied a new data management for identifying pathogen species based on mNGS data. We determined the thresholds for the unique reads and relative abundance required to identify the infectious pathogens. Potential pathogens of pulmonary infections in 15 patients were identified by mNGS. The comparison between mNGS and culture method resulted that the sensitivity and specificity were 100.0% (95% CI: 31.0-100.0%) and 76.5% (95% CI: 49.8-92.2%) for bacteria, 57.1% (95% CI: 20.2-88.2%) and 61.5% (95% CI: 32.2-84.9%) for fungi. The positive predictive value (PPV) (42.9% for bacteria, 44.4% for fungi) was much lower than negative predictive value (NPV) (100% for bacteria, 72.7% for fungi) in mNGS vs. culture method. The mNGS showed the highest specificity (100.0 and 94.1%) and PPV (100.0 and 75.0%) in the evaluation of fungi and MTBC respectively, when compared with histopathology method. The study indicated that mNGS of lung biopsy tissues can be used to detect the presence (or absence) of pulmonary pathogens in patients, with potential benefits in speed and sensitivity. However, accurate data management and interpretation of mNGS are required, and should be combined with observations of clinical manifestations and conventional laboratory-based diagnostic methods.

A sensitive signal-on photoelectrochemical sensor for tetracycline determination using visible-light-driven flower-like CN/BiOBr composites.

As a broad-spectrum antibiotic, tetracycline (TC) is widely used in agricultural purposes and human therapy. More attention is paid to TC as a serious threat to human health, including the fast spreading of antibiotic resistance gene and the serious toxicity to aquatic organisms. Therefore, the timely and accurate determination of TC residues is an urgent task to protect the safety of human. Herein, an effective and facile photoelectrochemical sensor platform based on carbon nitride/bismuth oxyhalide (CN/BiOBr) composites can be constructed for monitoring TC. The flower-like CN/BiOBr composites are prepared via a simple one-pot ethylene glycol-assisted solvothermal process with the addition of ionic liquid 1-hexadecyl-3-methylimidazolium bromide ([C16mim]Br). In view of matched energy band positions of CN and BiOBr, the addition of CN can reduce the recombination of photogenerated electron-hole pairs and improve the efficiency of visible light utilization, leading to enhancing photoelectrochemical response of BiOBr. Under light excitation, the photocurrent of CN/BiOBr composites is drastically improved, which is 6 times as much as that of pure BiOBr. Considering the superior photoelectrochemical performance, a photoelectrochemical sensor for monitoring TC has been developed, displaying linearly enhanced photocurrent with increasing the TC concentration. Two linear relationships received are from 8.0 to 4.0 × 102 ng mL-1, and 4.0 × 102 to 5.2 × 103 ng mL-1, respectively. The detection limit is 3.8 ng mL-1. The photoelectrochemical sensor exhibits a series of benefits including excellent stability, a wide linear range, a low detection limit and good anti-interference ability. Therefore, this work may offer great promises in providing a universal and efficient photoelectrochemical sensor for the tetracycline detection, and pave the way of constructing more materials used in photoelectrochemical detection field.

Fabrication of magnetically assembled aptasensing device for label-free determination of aflatoxin B1 based on EIS.

Aflatoxin B1 (AFB1), one of the most common mycotoxins in food matrixes, has been identified as the most toxic contaminant with mutagenic, teratogenic, immunosuppressive, and carcinogenic effects. In this work, a magnetically assembled aptasensing device has been designed for label-free determination of AFB1 by employing a disposable screen-printed carbon electrode (SPCE) covered with a designed polydimethylsiloxane (PDMS) film as the micro electrolytic cell. The magnetically controlled bio-probes were firstly prepared by immobilization of the thiolated aptamers on the Fe3O4@Au magnetic beads, which was rapidly assembled on the working electrode of SPCE within 10 s, by using a magnet placed at the opposite side. The PDMS film with a centered hole was covered on the SPCE surface to achieve a more practicable and flexible electrochemical measurement. In this effort, a label-free aptasensor for the sensitive and selective determination of AFB1 has been developed using electrochemical impedance spectroscopy upon the biorecognition between aptamers and the targets. The developed method had a wide linear range of 20 pg mL-1-50 ng mL-1 with a detection limit of 15 pg mL-1 (S/N = 3) and succeeded in spiked samples of peanuts. The developed aptasensing device shows fantastic application prospect with simple design, easy operation, low cost, and high sensitivity and selectivity characteristics. This sensing strategy represents a promising path toward routine quality control of food safety and creates the opportunity to develop facile aptasensing device for other targets.

NMI and IFP35 serve as proinflammatory DAMPs during cellular infection and injury.

Damage-associated molecular patterns (DAMP) trigger innate immune response and exacerbate inflammation to combat infection and cellular damage. Identifying DAMPs and revealing their functions are thus of crucial importance. Here we report that two molecules, N-myc and STAT interactor (NMI) and interferon-induced protein 35 (IFP35) act as DAMPs and are released by activated macrophages during lipopolysaccharide-induced septic shock or acetaminophen-induced liver injury. We show that extracellular NMI and IFP35 activate macrophages to release proinflammatory cytokines by activating nuclear factor-κB through the Toll-like receptor 4 pathway. In addition, the serum levels of NMI are increased in patients who succumbed to severe inflammation. NMI deficiency reduces inflammatory responses and mortality in mouse models of sepsis and liver injury. We therefore propose that extracellular NMI and IFP35 exacerbate inflammation as DAMPs, making them potential therapeutic targets for clinical intervention.Damage-associated molecular patterns (DAMP) are important mediators of innate immunity. Here the authors show that N-myc and STAT interactor (NMI) and interferon-induced protein 35 (IFP35) act as DAMPs to promote inflammation by activating macrophages via the Toll-like receptor 4 and NF-κB pathways.

Gold nanrods plasmon-enhanced photoelectrochemical aptasensing based on hematite/N-doped graphene films for ultrasensitive analysis of 17ß-estradiol.

It remains a vital task to establish ultrasensitive sensing interfaces for detection of target analytes to meet the demands of modern analysis. Herein, a highly sensitive turn-on photoelectrochemical (PEC) platform for trace 17ß-estradiol (E2) assay was developed based on Au nanrods (AuNRs) with surface plasmon resonance (SPR) properties induced signal amplification. Specifically, a ternary hybrid was prepared by integrating hematite (α-Fe2O3) nanocrystals and N-doped graphene (NG) with AuNRs, which further served as highly efficient photoactive species. Subsequently, a PEC sensing platform was fabricated based on the specific binding of E2 and its aptamer. On such a sensor, the capture of E2 molecules by aptamers led to increased photocurrent. This was attributed to that the specific recognition reaction between E2 and aptamer resulted in the conformational change of the aptamers and complete dissociation of some aptamers on the PEC sensing interface. It can be confirmed by the electrochemical impedance spectroscopy (EIS) results. This process decreased the steric hindrances between the electrode surface and solution and thus increased the photocurrent response. Under the optimal conditions, the as-prepared PEC aptasensor exhibited superb analytical performances for detection of E2 in the range from 1×10-15M to 1×10-9M with a detection limit of 3.3×10-16M. The aptasensor manifested outstanding selectivity towards E2 when other endocrine disrupting compounds with similar structure coexisted. Furthermore, the aptasensor was successfully applied for the determination of E2 in milk powder. The present strategy provides a potential way to boost the activity of photoactive materials and improve the sensitivity of PEC biosensor.

Health of US parents with and without disabilities.

BACKGROUND: The health of parents with disabilities is not well understood. Existing research has used small, non-representative samples. The lack of research using national representative data has hindered advocacy and policy-making efforts. OBJECTIVE: In the present study, we used nationally-representative data to examine the prevalence rates of chronic physical health conditions among parents with disabilities and compared them to parents without disabilities. METHODS: We analyzed pooled and linked data from the 2007-2011 Medical Expenditure Panel Survey and the corresponding National Health Interview Survey. We conducted logistic regression analyses to examine age-adjusted health differences of US parents with and without disabilities, controlling for covariates. Outcome measures included obesity, arthritis, asthma, cancer, heart disease, diabetes, emphysema, high cholesterol, hypertension, and stroke. RESULTS: After controlling for all model covariates and adjusting for age, parents with disabilities had significantly higher odds (aOR ranging from 1.69 to 4.82) of having each of the chronic conditions (P < 0.001). Parents with disabilities also have significant higher odds of having 2 conditions (aOR = 1.63), 3 conditions (aOR = 2.44), and 4 or more conditions (aOR = 5.56). CONCLUSIONS: Parents with disabilities have significantly poorer health than parents without disabilities.

High Prevalence of Hypervirulent Klebsiella pneumoniae Infection in China: Geographic Distribution, Clinical Characteristics, and Antimicrobial Resistance.

Hypervirulent Klebsiella pneumoniae (hvKP) is traditionally defined by hypermucoviscosity, but data based on genetic background are limited. Antimicrobial-resistant hvKP has been increasingly reported but has not yet been systematically studied. K. pneumoniae isolates from bloodstream infections, hospital-acquired pneumonia, and intra-abdominal infections were collected from 10 cities in China during February to July 2013. Clinical data were collected from medical records. All K. pneumoniae isolates were investigated by antimicrobial susceptibility testing, string test, extended-spectrum ß-lactamase (ESBL) gene detection, capsular serotypes, virulence gene profiles, and multilocus sequence typing. hvKP was defined by aerobactin detection. Of 230 K. pneumoniae isolates, 37.8% were hvKP. The prevalence of hvKP varied among different cities, with the highest rate in Wuhan (73.9%) and the lowest in Zhejiang (8.3%). Hypermucoviscosity and the presence of K1, K2, K20, and rmpA genes were strongly associated with hvKP (P < 0.001). A significantly higher incidence of liver abscess (P = 0.026), sepsis (P = 0.038), and invasive infections (P = 0.043) was caused by hvKP. Cancer (odds ratio [OR], 2.285) and diabetes mellitus (OR, 2.256) appeared to be independent variables associated with hvKP infections by multivariate analysis. Importantly, 12.6% of hvKP isolates produced ESBLs, and most of them carried blaCTX-M genes. Patients with neutropenia (37.5% versus 5.6%; P = 0.020), history of systemic steroid therapy (37.5% versus 5.6%; P = 0.020), and combination therapy (62.5% versus 16.7%; P = 0.009) were more likely to be infected with ESBL-producing hvKP. The prevalence of hvKP is high in China and has a varied geographic distribution. ESBL-producing hvKP is emerging, suggesting an urgent need to enhance clinical awareness, especially for immunocompromised patients receiving combination therapy.

CMTM7 knockdown increases tumorigenicity of human non-small cell lung cancer cells and EGFR-AKT signaling by reducing Rab5 activation.

The dysregulation of epidermal growth factor receptor (EGFR) signaling has been well documented to contribute to the progression of non-small cell lung cancer (NSCLC), the leading cause of cancer death in the world. EGF-stimulated EGFR activation induces receptor internalization and degradation, which plays an important role in EGFR signaling. This process is frequently deregulated in cancer cells, leading to enhanced EGFR levels and signaling. Our previous study on CMTM7 is only limited to a brief description of the relationship of overexpressed CMTM7 with EGFR-AKT signaling. The biological functions of endogenous CMTM7 and its molecular mechanism remained unclear. In this study, we show that the stable knockdown of CMTM7 augments the malignant potential of NSCLC cells and enhances EGFR-AKT signaling by decreasing EGFR internalization and degradation. Mechanistically, CMTM7 knockdown reduces the activation of Rab5, a protein known to be required for early endosome fusion. In NSCLC, the loss of CMTM7 would therefore serve to sustain aberrant EGFR-mediated oncogenic signaling. Together, our findings highlight the role of CMTM7 in the regulation of EGFR signaling in tumor cells, revealing CMTM7 as a novel molecule related to Rab5 activation.

Label-free photoelectrochemical immunosensor for neutrophil gelatinase-associated lipocalin based on the use of nanobodies.

Acute renal failure (ARF) represents a very important and potentially devastating disorder in clinical nephrology. Neutrophil gelatinase-associated lipocalin (NGAL) is an early biomarker for ARF in a wide range of different disease processes, which is frequently detected in clinical diagnosis. Herein, we present a label-free and sensitive photoelectrochemical (PEC) immunosensor for NGAL by utilizing a biotinylated anti-NGAL Nanobody (Nb) orientedly immobilized to streptavidin-coated cobalt 2,9,16,23-tetraaminophthalocyanine (CoPc)-sensitized TiO2 electrode. The Nb was biotinylated at the C-terminus, which is situated at the opposite site of the antigen binding region. Using highly oriented Nb as receptor molecules, a label-free PEC immunosensor for NGAL was developed by monitoring the changes in the photocurrent signals of the electrode resulting from immunoreaction. Immobilization of Nb to streptavidin-coated CoPc-sensitized TiO2 electrode surface provides high binding capacity to NGAL; thus, it can lead to a high sensitivity. The limit of detection (LOD) of the proposed immunosensor has been significantly lowered to 0.6 pg mL(-1). This proposed immunosensor reveals high specificity to detect NGAL, with acceptable intra-assay precision and excellent stability. In addition, the present work provides a new approach to design Nb-based PEC immunosensor and increases versatility of Nbs.

Synthesis of fluorescent dye-doped silica nanoparticles for target-cell-specific delivery and intracellular microRNA imaging.

MicroRNA (miRNA) is found to be up-regulated in many kinds of cancer and therefore is classified as an oncomiR. Herein, we design a multifunctional fluorescent nanoprobe (FSiNP-AS/MB) with the AS1411 aptamer and a molecular beacon (MB) co-immobilized on the surface of the fluorescent dye-doped silica nanoparticles (FSiNPs) for target-cell-specific delivery and intracellular miRNA imaging. The FSiNPs were prepared by a facile reverse microemulsion method from tetraethoxysilane and silane derivatized coumarin that was previously synthesized by click chemistry. The as-prepared FSiNPs possess uniform size distribution, good optical stability and biocompatibility. In addition, there is a remarkable affinity interaction between the AS1411 aptamer and the nucleolin protein on the cancer cell surface. Thus, a target-cell-specific delivery system by the FSiNP-AS/MB is proposed for effectively transferring a MB into the cancer cells to recognize the target miRNA. Using miRNA-21 in MCF-7 cells (a human breast cancer cell line) as a model, the proposed multifunctional nanosystems not only allow target-cell-specific delivery with the binding affinity of AS1411, but also can track simultaneously the transfected cells and detect intracellular miRNA in situ. The proposed multifunctional nanosystems are a promising platform for a highly sensitive luminescent nonviral vector in biomedical and clinical research.

Label-free fluorescence detection of DNA methylation and methyltransferase activity based on restriction endonuclease HpaII and exonuclease III.

Strategies to detect the methylation of site specific DNA and assay of M.SssI methyltransferase (M.SssI MTase) activity are important in determining human cancers due to aberrant methylation linked to cancer initiation and progression. Herein, we report a label-free fluorescence detection method for DNA methylation and MTase activity based on restriction endonuclease HpaII and exonuclease III (Exo III). A label-free probe DNA was designed, which hybridized with target DNA (one 32-mer DNA from the exon 8 promoter region of the Homo sapiens p53 gene) to form double stranded DNA (dsDNA). Upon the cleavage action of HpaII and degradation reaction of Exo III, dsDNA changed to single stranded DNA (ssDNA) and the fluorescence intensity of thiazole orange (TO) is weak. After the resulting dsDNA was methylated by M.SssI MTase, the action of HpaII and Exo III was prevented, then TO intercalates into the dsDNA and emits strong fluorescence. This method can determine DNA methylation at the site of CpG and distinguish a one-base mismatched target sequence. The fluorescence intensity has a linear relationship with M.SssI MTase activities in the range of 1-10 U mL(-1) with a detection limit of 0.16 U mL(-1) in terms of 3 times deviation of the blank sample. The methylation of DNA by a hydroxyl radical triggered by DMSO and CH3CHO was also measured. These results show that the proposed method can specifically and selectively detect DNA methylation and M.SssI MTase activity. Human serum has no obvious effects on the assay performance, indicating that the method has great potential for further application in complex samples.