Target proteins-regulated DNA nanomachine-electroactive substance complexes enable separation-free electrochemical detection of clinical exosome.

Simple and reliable profiling of tumor-derived exosomes (TDEs) holds significant promise for the early detection of cancer. Nonetheless, this remains challenging owing to the substantial heterogeneity and low concentration of TDEs. Herein, we devised an accurate and highly sensitive electrochemical sensing strategy for TDEs via simultaneously targeting exosomal mucin 1 (MUC1) and programmed cell death ligand 1 (PD-L1). This approach employs high-affinity aptamers as specific recognition elements, utilizes rolling circle amplification and DNA nanospheres as effective bridges and signal amplifiers, and leverages methylene blue (MB) and doxorubicin (DOX) as robust signal reporters. The crux of this separation- and label-free method is the specific response of MB and DOX to G-quadruplex structures and DNA nanospheres, respectively. Quantifying TDEs using this strategy enabled precise discrimination of lung cancer patients (n = 25) from healthy donors (n = 12), showing 100% specificity (12/12), 92% sensitivity (23/25), and an overall accuracy of 94.6% (35/37), with an area under the receiver operating characteristic curve (AUC) of 0.97. Furthermore, the assay results strongly correlated with findings from computerized tomography and pathological analyses. Our approach could facilitate the early diagnosis of lung cancer through TDEs-based liquid biopsy.

Nucleic Acid and Nanomaterial Synergistic Amplification Enables Dual Targets of Ultrasensitive Fluorescence Quantification to Improve the Efficacy of Clinical Tuberculosis Diagnosis.

Interferon-γ (IFN-γ) release assays (IGRAs) are constrained by the limited diagnostic performance of a single indicator and the excessive Mycobacterium tuberculosis (Mtb) antigen stimulation time. This study presents a simultaneous, homogeneous, rapid, and ultrasensitive fluorescence quantification strategy for IFN-γ and IFN-γ-induced protein 10 (IP-10). This method relies on the high-affinity binding of aptamers to IFN-γ and IP-10, the enzyme-free catalytic hairpin assembly reaction, and the heightened sensitivity of CdTe quantum dots to Ag+ and hairpin structure C-Ag+-C and carbon dots to Hg2+ and hairpin structure T-Hg2+-T. Under optimized conditions, the selectivity of IFN-γ and IP-10 was excellent, with a linear range spanning from 1 to 100 ag/mL and low limits of detection of 0.3 and 0.5 ag/mL, respectively. Clinical practicality was confirmed through testing of 57 clinical samples. The dual-indicator combination detection showed 92.8% specificity and 93.1% sensitivity, with an area under the curve of 0.899, representing an improvement over the single-indicator approach. The Mtb antigen stimulation time was reduced to 8 h for 6/7 clinical samples. These findings underscore the potential of our approach to enhance the efficiency and performance of a tuberculosis (TB) clinical diagnosis.

Single-Cell Liquid Biopsy of Lung Cancer: Ultra-Simplified Efficient Enrichment of Circulating Tumor Cells and Hand-Held Fluorometer Portable Testing.

Herein, we propose a paper-based laboratory via enzyme-free nucleic acid amplification and nanomaterial-assisted cation exchange reactions (CERs) assisted single-cell-level analysis (PLACS). This method allowed for the rapid detection of mucin 1 and trace circulating tumor cells (CTCs) in the peripheral blood of lung cancer patients. Initially, an independently developed method requiring one centrifuge, two reagents (lymphocyte separation solution and erythrocyte lysate), and a three-step, 45 min sample pretreatment was employed. The core of the detection approach consisted of two competitive selective identifications: copper sulfide nanoparticles (CuS NPs) to C-Ag+-C and Ag+, and dual quantum dots (QDs) to Cu2+ and CuS NPs. To facilitate multimodal point-of-care testing (POCT), we integrated solution visualization, test strip length reading, and a self-developed hand-held fluorometer readout. These methods were detectable down to ag/mL of mucin 1 concentration and the single-cell level. Forty-seven clinical samples were assayed by fluorometer, yielding 94% (30/32) sensitivity and 100% (15/15) specificity with an area under the curve (AUC) of 0.945. Nine and 15 samples were retested by a test strip and hand-held fluorometer, respectively, with an AUC of 0.95. All test results were consistent with the clinical imaging and the folate receptor (FR)-PCR kit findings, supporting its potential in early diagnosis and postoperative monitoring.

The importance of biofilm contamination control for dental unit waterlines: a multicenter assessment of the microbiota diversity of biofilm in dental unit waterlines.

Background: The biofilm formation in Dental Unit Waterlines (DUWLs) could become an important cause of infection during dental care, which could put immunocompromised individuals at risk of cross-infection. The aim of this study was to characterize the microbial communities of biofilms among DUWLs using high-throughput sequencing technology. Methods: Twenty-nine biofilm samples were obtained from 24 dental chair units at 5 hospitals and 2 dental clinics. The genomic DNA of the samples was extracted, then 16S rDNA and ITS2 gene were amplified and sequenced. Alpha-diversity and Beta-diversity were calculated with QIIME2 and the Kruskal - Wallis H-test was adopted for statistical analysis. Results: Microbial communities with a high diversity of bacteria (377 genera) and fungi (83 genera) were detected in the biofilm samples. The dominant phylum of bacteria was Proteobacteria (93.27%) and that of fungi was Basidiomycota (68.15%). Potential human pathogens were detected including 7 genera of bacteria (Pseudomonas, Stenotrophomonas, Hafnia-Obesumbacterium, Burkholderia-Caballeronia-Paraburkholderia, Ralstonia, Enterobacter, Klebsiella) and 6 genera of fungi (Malassezia, Candida, Alternaria, Cryptococcus, Rhodotorula, Rhinocladiella). Conclusions: This multicenter assessment revealed the infectious risk during dental care. It emphasized the importance of biofilm control due to biofilm accumulation and multiple kinds of opportunistic pathogens in DUWLs.

Biomolecules-mediated electrochemical signals of Cu2+: Y-DNA nanomachines enable homogeneous rapid one-step assay of lung cancer circulating tumor cells.

This study presents a straightforward efficient technique for extracting circulating tumor cells (CTCs) and a rapid one-step electrochemical method (45 min) for detecting lung cancer A549 cells based on the specific recognition of mucin 1 using aptamers and the modulation of Cu2+ electrochemical signals by biomolecules. The CTCs separation and enrichment process can be completed within 45 min using lymphocyte separation solution (LSS), erythrocyte lysis solution (ELS), and three centrifugations. Besides, the influence of various biomolecules on Cu2+ electrochemical signals is comprehensively discussed, with DNA nanospheres selected as the medium. Three single-stranded DNA sequences were hybridized to form Y-shaped DNA (Y-DNA), creating DNA nanospheres. Upon specific capture of mucin 1 by the aptamer, most DNA nanospheres could form complexes with Cu2+ (DNA nanosphere-Cu2+), significantly reducing the concentration of free Cu2+. Our approach yielded the limit of detection (LOD) of 2 ag/mL for mucin 1 and 1 cell/mL for A549 cells. 39 clinical blood samples were used for further validation, yielding results closely correlated with pathological, computed tomography (CT) scan findings and folate receptor-polymerase chain reaction (FR-PCR) kits. The receiver operating characteristic (ROC) curve displayed an area under the curve (AUC) value of 0.960, demonstrating 100% specificity and 93.1% sensitivity for the assay. Taken together, our findings indicate that this straightforward and efficient pretreatment and rapid, highly sensitive electrochemical assay holds great promise for liquid biopsy-based tumor detection using CTCs.

Controllable self-assembled DNA nanomachine enable homogeneous rapid electrochemical one-pot assay of lung cancer circulating tumor cells.

A homogeneous rapid (45 min) one-pot electrochemical (EC) aptasensor was established to quantitatively detect circulating tumor cells (CTCs) in lung cancer patients using mucin 1 as a marker. The core of this study is that the three single-stranded DNA (Y1, Y2, and Y3) could be hybridized to form Y-shaped DNA (Y-DNA) and further self-assemble to form DNA nanosphere. The aptamer of mucin 1 could be complementary and paired with Y1, thus disrupting the conformation of the DNA nanosphere. When mucin 1 was present, the aptamer combined specifically with mucin 1, thus preserving the DNA nanosphere structure. Methylene blue (MB) acted as a signal reporter, which could be embedded between two base pairs in the DNA nanosphere to form a DNA nanosphere-MB complex, reducing free MB and resulting in a lower electrochemical signal. The results demonstrated that the linear ranges for mucin 1 and A549 cells were 1 ag/mL-1 fg/mL and 1-100 cells/mL, respectively, with minimum detectable concentrations were 1 ag/mL and 1 cell/mL, respectively. The quantitative analysis of CTCs in 44 clinical blood samples was performed, and the results were consistent with the computerized tomography (CT) images, pathological findings and folate receptor-polymerase chain reaction (FR-PCR) kits. The receiver operating characteristic (ROC) curve exhibited an area under the curve (AUC) value of 0.970. The assay revealed 100% specificity and 94.1% sensitivity. It is believed that this electrochemical aptasensor could provide a new approach to detect CTCs.

Simplified Rapid Enrichment of CTCs and Selective Recognition Prereduction Enable a Homogeneous ICP-MS Liquid Biopsy Strategy of Lung Cancer.

The effective enrichment and hypersensitivity analysis of circulating tumor cells (CTCs) in clinical whole blood samples are highly significant for clinical tumor liquid biopsy. In this study, we established an easy operation and affordable CTCs extraction technique while simultaneously performing the homogeneous inductively coupled plasma mass spectrometry (ICP-MS) determination of CTCs in lung cancer clinical samples based on selective recognition reactions and prereduction phenomena. Our strategy allowed for the pretreatment of whole blood samples in less than 45 min after step-by-step centrifugation, which only required lymphocyte separation solution and erythrocyte lysate. Furthermore, a three-stage signal amplification system consisting of catalytic hairpin assembly (CHA), selective recognition for C-Ag+-C structures and Ag+ of copper sulfide nanoparticles (CuS NPs), and prereduction of Hg2+ through ascorbic acid (AA) was constructed by using mucin 1 as the CTCs marker and the aptamer for identification probes. In optimal conditions, the detection limits of ICP-MS were as low as 0.3 ag/mL for mucin 1 and 0.25 cells/mL for A549 cells. This method analyzed CTCs in 58 clinical samples quantitatively, and the results were consistent with clinical CT images and pathological findings. The area under the curve (AUC) value of the receiver operating characteristic (ROC) curve was 0.957, which provided a specificity of 100% and a sensitivity of 91.5% for the assay. Therefore, the simplicity of the extraction method, the accessibility, and the high sensitivity of the assay method make the strategies attractive for clinical CTCs testing applications.

Transcriptomic analysis reveals the rareness of genetic assimilation of gene expression in environmental adaptations.

Genetic assimilation is the evolutionary process by which an environmentally induced phenotype becomes genetically encoded and constitutive. Genetic assimilation has been proposed as a concluding step in environmental adaptation, but its prevalence has not been systematically investigated. Analyzing transcriptomic data collected upon reciprocal transplant, we address this question in the experimental evolution, domestication, or natural evolution of seven diverse species. We find that genetic assimilation of environment-induced gene expression is the exception rather than the rule and that substantially more genes retain than lose their expression plasticity upon organismal adaptations to new environments. The probability of genetic assimilation of gene expression decreases with the expression level and number of transcription factors controlling the gene, suggesting that genetic assimilation results primarily from passive losses of gene regulations that are not mutationally robust. Hence, for gene expression, our findings argue against the purported generality or importance of genetic assimilation to environmental adaptation.

Homogeneous Dual Fluorescence Count of CD4 in Clinical HIV-Positive Samples via Parallel Catalytic Hairpin Assembly and Multiple Recognitions.

Regularly measuring the level of CD4+ cells is necessary for monitoring progression and predicting prognosis in patients suffering from an infection with the human immunodeficiency virus (HIV). However, the current flow cytometry standard detection method is expensive and complicated. A parallel catalytic hairpin assembly (CHA)-assisted fluorescent aptasensor is reported for homogeneous CD4 count by targeting the CD4 protein expressed on the membrane of CD4+ cells. Detection was achieved using CdTe quantum dots (QDs) and methylene blue (MB) as signal reporters. CdTe QDs distinguished CHA-assisted release of Ag+ and C-Ag+-C and MB that has differentiated cytosine (C)-rich single-stranded DNA (ssDNA) and C-Ag+-C, generating changes in fluorescence intensity. With the assistance of the CHA strategy and luminescent nanomaterials, this method reached limits of detection of 0.03 fg/mL for the CD4 protein and 0.3 cells/mL for CD4+ cells with linear ranges of 0.1 to 100 fg/mL and 1 to 1000 cells/mL, respectively. The method was validated in 50 clinical whole blood samples consisting of 30 HIV-positive patients, 10 healthy volunteers, and 10 patients with cancer or other chronic infections. The findings from this method were in good agreement with the data from clinical flow cytometry. Due to its sensitivity, affordability, and ease of operation, the current method has demonstrated great potential for routine CD4 counts for the management of HIV, especially in communities and remote areas.

Target Enzyme-Triggered Click Chemistry and Hybridization Chain Reaction for Fluorescence Nonculture Homogeneous Analysis of E. coli in Bloodstream Infections.

Escherichia coli is the major pathogen that causes bloodstream infections (BSI). It is critical to develop nonculture identification methods which can meet the urgent need of clinical diagnosis and treatment. In this study, we reported a homogeneous fluorescence E. coli analysis system using ß-galactosidase (ß-Gal) as the biomarker and double-stranded DNA-templated copper nanoparticles (dsDNA-Cu NPs) as the signal output. The product of the enzymatic hydrolysis reaction, p-aminophenol (PAP), could reduce Cu2+ to Cu+, triggering the alkyne-azido cycloaddition reaction (CuAAC). Subsequently, the hybrid chain reaction (HCR) was initiated, producing the dsDNA template used to generate Cu NPs in situ. The system achieved a wide linear range for ß-Gal and E. coli 1-104 mU/L and 10-2-10 colony-forming unit (CFU)/mL, and a detection limit of 0.3 mU/L and 0.003 CFU/mL, respectively. 65 samples (45 blood and 20 urine) were collected to evaluate the clinical practicality. The results demonstrated remarkable area under the curve (AUC) values of 0.95 and 0.916 from uncultured urine and blood, respectively. It had 100% specificity and 83.3% sensitivity. The whole duration of the strategy was 3.5 h, which significantly reduced the turnaround time (TAT) and facilitated early BSI diagnosis to improve patients' prognosis. Our work had the potential to be an alternative to culture-based methods in clinics.

Nanoarchitectonics-Assisted Simultaneous Fluorescence Detection of Urinary Dual miRNAs for Noninvasive Diagnosis of Prostate Cancer.

Herein, we report a fluorescence strategy for the homogeneous and simultaneous analysis of urine miRNA-375 and miRNA-148a. The target miRNAs in urine bonded the devised dumbbell-shaped "C-Ag+-C" and "T-Hg2+-T" hairpin structures that could trigger cascade enzyme-free amplification. Then, the fluorescent CdTe quantum dots (QDs) and carbon dots (CDs) could selectively recognize Ag+ and Hg2+, to quantify the dual miRNAs concurrently. Under optimized conditions, the linear range was from 0.1 to 1000 fM and the limits of detection (LOD) for dual miRNAs reached 30 and 25 aM, respectively. The practicality was further evaluated with 45 clinical urine samples including prostate cancer (PC) and other patients, and the results were consistent with the clinical polymerase chain reaction (PCR) kit and ultrasonic and pathological findings. The receiver operating characteristic (ROC) curve analysis showed that the estimates of the area under the curve (AUC) were 0.739 for the serum prostate-specific antigen (PSA) and 0.941 for miRNA-375 and 0.946 for miRNA-148a. The sensitivity and specificity reached 75 and 100% for miRNA-375 and 71 and 94% for miRNA-148a, respectively, which was better than serum PSA. This strategy constructed a reliable system for dual miRNA detection in urine samples and proposed new insights into the rapid and noninvasive diagnosis of PC.

Metal ions-regulated chemical vapor generation of Hg2+:mechanism and application in miniaturized point discharge atomic emission spectrometry assay of oxalate in clinical urolithiasis samples.

It is well known that the coexisting metal ions could significantly influence the atomic spectroscopy (AS) analysis. In this work, a cation-modulated mercury ions (Hg2+) strategy via chemical vapor generation (CVG) was developed for oxalate assay due to the phenomenon that the Ag + can significantly reduce the Hg2+ signal. The regulation effect was studied in depth via experimental investigations. Since Ag + can be reduced to silver nanoparticles (Ag NPs) by reductant SnCl2, the decrease of the Hg2+ signal is attributed to the formation of a silver-mercury (Ag-Hg) amalgam. Due to the oxalate can react with Ag + to generate Ag2C2O4, which can reduce the generation of Ag-Hg amalgam, a portable and low-power point discharge chemical vapor generation atomic emission spectrometry (PD-CVG-AES) system was constructed to quantify the content of oxalate via monitoring the signal of Hg2+. Under optimal conditions, the limit of detection (LOD) was as low as 40 nM in the range of 0.1-10 µM for oxalate assay, while exhibiting good specificity. This method was applied to quantitative oxalate in 50 clinical urine samples of urinary stones patients. The levels of oxalate detected in clinical samples were consistent with clinical imaging results, which is promising for point-of-care testing in clinical diagnosis.

Sensitive Urine Immunoassay for Visualization of Lipoarabinomannan for Noninvasive Tuberculosis Diagnosis.

Lipoarabinomannan (LAM) is a prospective noninvasive biomarker for tuberculosis (TB) diagnosis. Here, we report a visual immunoassay of high sensitivity for detecting LAM in urine samples toward TB diagnosis. This method uses a DNA-linked immunosorbent of LAM, followed by a transduction cascade into amplified visual signals using quantum dots (QDs) and calcein reaction with Cu2+ and copper nanoparticles (Cu NPs). The limit of detection (LOD) for LAM in the urine reaches 2.5 fg/mL and 25 fg/mL using a fluorometer and length readouts on strips, respectively, demonstrating an ultrahigh sensitivity. The clinical validation of the proposed assay was performed with 147 HIV-negative clinical urine specimens. The results show the sensitivity of test is 94.1% (16/17) for confirmed TB (culture-positive) and 85% (51/60) for unconfirmed TB (clinical diagnosis without positive culture results), respectively, when the test cutoff value is 40 fg/mL for TB. Its specificity is 89.2% (25/28) in non-TB and nontuberculous mycobacterial patients. The area under the curve (AUC) was 0.86 when controls were non-TB and LTBI patients, while the AUC was 0.92 when controls were only non-TB patients. This highly sensitive visual immunoassay of LAM has shown potential for noninvasive diagnosis of TB using urine samples.

Rapid binary visual detection of oxalate in urine samples of urolithiasis patients via competitive recognition and distance reading test strips.

Urolithiasis is a common disease with wide ranging effects, with oxalate stones being the most prevalent type. Existing clinical diagnostic methods rely on complex instruments and professionals, are difficult to distinguish between stone types, and have insufficient sensitivity. Moreover, high-sensitivity point-of-care testing (POCT) methods remain scarce. We constructed a rapid homogeneous dual fluorescence and binary visualization analysis system to diagnose oxalate urolithiasis because oxalate can efficiently reduce Cu2+ to Cu+, which can be selectively competitively recognized by both calcein and cadmium telluride quantum dots (CdTe QDs). Under optimized conditions, the system exhibited high sensitivity to oxalate ranging from 10 pM to 10 nM within 3 min. Following that, visualized test strips of calcein and QDs were generated by inkjet printing; oxalate concentrations as low as 10 nM can be easily identified by reading the quenching distance on the strip. We then analyzed 66 clinical urine samples: 11 healthy, 10 oxalate-negative, and 45 oxalate-positive samples. The fluorescence and visual mode results were highly consistent with clinical computed tomography (CT) images and clinical diagnostics. Therefore, our analysis strategy has the potential to use POCT for the assessment of oxalate urolithiasis.

Three-fluorescence sensor for minute-time scale low-cost analysis of urinary oxalate in urolithiasis metabolic assessment.

Since oxalate plays an important role in the metabolic assessment of urolithiasis, there is need for convenient and efficient methods for oxalate detection. Herein, we report a three-signal fluorescence strategy for oxalate analysis based on the ability of oxalate to reduce Cu2+ to Cu+, and the ability of pyrophosphate-cerium coordination polymeric networks (PPi-Ce CPNs), cadmium telluride quantum dots (CdTe QDs), and N-Methyl Mesoporphyrin (NMM) to selectively detect Cu2+ and Cu+. The detection range was 100 nM to 1 mM, the turnaround time was 6 min, while the limits of detections for PPi-Ce CPNs, QDs and NMM as reporters were 25 nM, 10 nM and 40 nM, respectively. Visual detection of oxalate relied on color change in the solution, which could be observed using the naked eye. The fluorescent system was used for oxalate analysis in 44 urine samples (32 calcium oxalate stone patients, 12 controls without urolithiasis), and the results were consistent with clinical diagnosis and imaging data. Moreover, the visual system was used to analyze 8 urine samples (4 patients and 4 controls), and showed good consistency with clinical diagnosis and computed tomography imaging results. These findings suggest that the method has potential application for the metabolic assessment of urolithiasis.

Prevalence of dental anxiety among adults in China: a Meta-analysis.

OBJECTIVES: This study aimed to systematically evaluate the prevalence of dental anxiety in Chinese adults and to provide references for decision making on oral healthcare. METHODS: We searched PubMed, Web of Science, Ebsco, Embase, The Cochrane Library, WanFang Data, CNKI, and VIP database to collect cross-sectional studies on dental anxiety in Chinese adults from the establishment of the databases to 30 September 2022. After literature screening, data extraction, and evaluation of the risk of bias in the included studies by two researchers independently, R 4.0.4 software was used to perform a Meta-analysis. RESULTS: A total of 39 studies were included, including 24 309 subjects. Meta-analysis showed that the prevalence of dental anxiety in Chinese adults was 35.39% [95%CI (31.31%, 40.01%)]. Subgroup analysis showed that the prevalence rates of male and female adults were 32.92% and 44.78%, respectively. The prevalence rates of adults aged 16-39,40-59, ≥60 were 49.37%, 47.13%, and 37.41%, respectively. The prevalence rates of mild, moderate, and severe patients were 13.81%, 15.15%, and 9.24%, respectively. The prevalence rates of adults with elementary school and below, middle school, and university and above education levels were 33.81%, 35.84%, and 36.24%, respectively. The prevalence rates were 39.45% and 45.90% in adults with and without dental-treatment history, respectively. The prevalence rates of adults surveyed in dental and non-dental clinics were 27.10% and 39.31%, respectively. CONCLUSIONS: The prevalence of dental anxiety in Chinese adults was relatively high, primarily moderate anxiety, and it was more likely to occur in women, young people, and groups with no history of dental treatment. Early intervention should be performed for adults with dental anxiety to improve their awareness of oral healthcare and treatment compliance and thus to promote the oral-health level of adults in China.

Prevalence of periodontal disease in pregnancy: A systematic review and meta-analysis.

OBJECTIVES: The purpose of this study was to systematically assess the epidemic trend of periodontal disease in pregnancy. DATA: Eligibility criteria comprised studies that reported periodontitis and the periodontal indicators of BOP (+) or CAL≥4 mm or PD≥4 mm among pregnant women. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were applied where applicable. Risk of bias was assessed using the Critical Appraisal Checklist for prevalence studies proposed by The Joanna Briggs Institute (JBI). Meta-analyses were conducted to estimate the pooled effect measures. Q-statistic, I2 statistic, subgroup and sensitivity analyses assessed study heterogeneity. SOURCES: Electronic search of articles was conducted using PubMed, Web of Science, EMBASE, Scopus, and Ovid from January 2000 to January 2022. RESULTS: A total of 20 studies were included in the meta-analysis. The prevalence of periodontitis among pregnancy was 40% (95% Confidence Interval (CI): [0.15, 1.00]). The prevalence rates were 67% (CI [0.56, 0.80]), 42% (CI [0.27, 0.57]) and 24% (CI [0.12, 0.37]) for BOP (+), PD≥4 mm and CAL≥4 mm respectively. Regarding subgroup meta-analyses, the prevalence rates of BOP (+) and PD≥4 mm presented a gradual increase throughout pregnancy, while the highest prevalence rate of CAL≥4 mm was in the 2nd trimester. CONCLUSIONS: It was observed a high prevalence of periodontal disease in pregnancy. However, heterogeneity was high among included studies. More high-quality epidemiologic investigations on periodontal disease in pregnancy are still needed. CLINICAL SIGNIFICANCE: Periodontal disease in pregnancy is highly prevalent which results in a reduced quality of life, frequent systemic pathologies and adverse pregnancy outcomes. Given the unhealthy consequences, public health impact, and expansive disease burden, it is worthwhile to investigate more aspects of periodontal disease during pregnancy.

A microplasma optical emission spectrometry pen for point-of-care diagnosis of child blood lead.

Blood lead levels (BLL) of children have attracted considerable attention due to their putative impact on intelligence decline. However, most methods used for the determination of blood lead typically require expensive, bulky, high power and gas consuming instrumentation, limiting their application for a point-of-care diagnosis. Herein we report the development and testing of a portable ballpoint discharge microplasma optical emission spectrometer (BD-OES pen) device having the potential to fill this needed measurement capability. The BD-OES pen utilizes a compact ballpoint-pen format integrating point-discharge microplasma, which permits the determination of child BLL requiring no more than 100 µL blood while providing high specificity, sensitivity and satisfactory limit of detection (0.73 µg L-1). The handheld BD-OES pen is successfully used to diagnose BLL of 16 asymptomatic children on-site, two of whom had excessive the normal BLL. The pen may aid the on-site and rapid diagnosis of childhood BLL, particularly in low-income areas.

Simultaneous Homogeneous Fluorescence Detection of AFP and GPC3 in Hepatocellular Carcinoma Clinical Samples Assisted by Enzyme-Free Catalytic Hairpin Assembly.

Simultaneous sensitive and cost-effective detection of multiple tumor markers has shown great potential for cancer diagnostics. Herein, we reported a simple enzyme-free parallel catalytic hairpin assembly (CHA) amplification strategy with N-methyl mesoporphyrin IX (NMM) and quantum dots (QDs) as signal reporters for the homogeneous fluorescent simultaneous detection of alpha-fetoprotein (AFP) and glypican-3 (GPC3). Upon selective binding, the released single-stranded DNA (ssDNA) from the two-aptamer double-stranded DNA (dsDNA) probes triggers CHA amplification, further releasing the G-quadruplex sequence and Ag+ from the C-Ag+-C structures at the same time. Then, NMM and CdTe QDs selectively recognize G-quadruplex and Ag+, respectively. Under optimized conditions, limits of detections (LODs) as low as 3 fg/mL for AFP and 0.25 fg/mL for GPC3 were achieved using fluorescence readout. Using color- and distance-based visual readouts, an LOD of 1 fg/mL for GPC3 was reached. This method was applied to quantitatively analyze AFP and GPC3 in 41 clinical serum samples of hepatocellular carcinoma (HCC) patients. The quantitative test results for AFP and GPC3 were consistent with those obtained using the electrochemiluminescence immunoassay (ECL-IA) clinical kit and correlated with radiological and pathological findings. The results of clinical tests demonstrated the potential of GPC3 as a tumor biomarker, and we propose a cut-off value of 2 ng/mL GPC3 for HCC.

Fluorescence Aptasensor of Tuberculosis Interferon-γ in Clinical Samples Regulated by Steric Hindrance and Selective Identification.

Although there are many interferon gamma (IFN-γ)-based tools for tuberculosis (TB) diagnosis, they are less sensitive and laborious. Here, we developed an IFN-γ aptasensor using pyrophosphate-cerium coordination polymeric nanoparticles (PPi-Ce CPNs) as signal reporters and a double-stranded DNA as a probe. The sensor was realized by sterically regulating the polymerization elongation of terminal deoxynucleotidyl transferase (TdT) and the selective recognition reaction of PPi-Ce CPNs. This method employs PPi-Ce CPNs to selectively identify Cu2+ and polyT-templated copper nanoparticles (Cu NPs), as well as a TdT-assisted amplification technique. Our data showed that under optimized experimental conditions, a limit of detection of as low as 0.25 fg/mL was achieved, with a linear range of 1-100 fg/mL, and a good target protein specificity. The detection sensitivity was an order of magnitude higher than that observed with Cu NPs when used as signal reporters. This IFN-γ quantification technique was further validated in clinical samples using 57 clinical TB patients (22 negative and 35 positive). Our findings agreed with those from enzyme-linked immunosorbent assay, GeneXpert MTB/rifampin assay, and polymerase chain reaction detection of TB-DNA and those from clinical imaging techniques. Therefore, our analytical system may provide an additional and more sensitive tool for the early diagnosis of TB.