Biomimetic and multifunctional nanocomposites for precision fungi theranostics.

Fungi infection is a serious threat to public health, but an effective antifungal strategy remains a challenge. Herein, a biomimetic nanocomposite with multifunctionalities, including fungi diagnosis, antifungal adhesion, precise fungi elimination, and cytokine sequestration, is constructed for battling Candida albicans (C. albicans) infection. By screening a range of cells, we find that the polarized macrophage cells have the strongest binding tendency toward C. albicans. Thus, their membranes were exfoliated to camouflage UCNPs and then decorated with photosensitizers (methylene blue, MB) and DNA sensing elements. The resulting nanocomposite can tightly bind to fungal surfaces, promote DNA recognition, and squeeze pro-inflammatory cytokines to relieve inflammation. Consequently, this nanocomposite can detect C. albicans with enhanced sensitivity and precisely eliminate fungal cells through photodynamic therapy with minimal phototoxicity because of its switchable fluorescence behavior. The developed nanocomposite with good biocompatibility achieves a satisfactory diagnostic and therapeutic effect in a C. albicans-infected mouse model, which offers a unique approach to fight fungi infection.

A Controlled Clinical Study of Accelerated High-Dose Theta Burst Stimulation in Patients with Obsessive-Compulsive Disorder.

Background: Obsessive-compulsive disorder (OCD) is frequently treated using a combination of counseling, drugs, and, more recently various transcranial stimulation protocols, but all require several weeks to months for clinically significant improvement, so there is a need for treatments with faster onset. This study investigated whether an accelerated high-dose theta burst stimulation (ahTBS) protocol significantly improves the efficacy of OCD compared to traditional 1-Hz repetitive transcranial magnetic stimulation (rTMS) in the routine clinical setting. Method: Forty-five patients with OCD were randomized into two groups and treated with ahTBS or 1-Hz rTMS for 5 days. Patients were assessed at baseline at the end of treatment using the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS). Results: After 5 days of treatment, there was a significant decrease in Y-BOCS scores in both groups (p < 0.001), and the difference between the two groups was not statistically significant (group × time interaction, F = 1.90, p=0.18). There was also no statistically significant difference in other secondary outcome indicators, including depression, anxiety symptoms, and response rate. However, the ahTBS group had a greater trend in response rate. Neuropsychological testing showed no negative cognitive side effects of either treatment. Conclusion: Accelerated high-dose TBS is as safe and has comparable short-term efficacy to traditional 1-Hz rTMS for the clinical treatment of OCD. Further research is needed to explore optimal ahTBS parameters, validate the utility of this treatment modality, and identify factors predictive of rapid clinical response to guide clinical decision-making. This trial is registered with NCT05221632.

Exposure to metal mixtures and young children's growth and development: A biomonitoring-based study in Eastern China.

Exposure to metal mixtures may affect children's health but the conclusions are controversial. We aimed to investigate the associations of metal mixture exposure with children's physical and behavioral development. 15 metals were detected in the urine samples of 278 preschoolers aged 3-6 years from eastern China. Multiple linear models and restricted cubic splines were used to evaluate dose-response relationships between single metal and children's physical and behavioral development. The Bayesian Kernel Machine Regression (BKMR) models, the weighted quantile sum (WQS) models and Quantile G-Computation were applied to evaluate the joint effects of metal mixtures. The results showed that arsenic (As) was negatively associated with z score of height for age (HAZ) in individual-metal models [ß (95%CI): - 0.22 (-0.38, -0.06), P = 0.006]. Concerning children's behavioral development, multiple-metal models demonstrated a negative association with strontium (Sr) [ß (95%CI): - 0.82 (-1.38, -0.26), P = 0.004], and a positive association with tin (Sn) [ß (95%CI): 0.69 (0.16, 1.21), P = 0.010]. Notably, these associations remained significant or suggestive even after adjustments for multiple tests, sensitivity analyses, and application of different statistical models, including BKMR, WQS, and Quantile G-Computation. Furthermore, the study identified a negative joint effect of the metal mixture on HAZ, as demonstrated by BKMR and Quantile G-Computation models, with As playing an irreplaceable role in this observed impact. In summary, exposure to As appears to have adverse effects on HAZ, while exposure to Sn may hinder children's behavioral development. Conversely, exposure to Sr may have a protective effect on children's behavioral development. Additionally, the combined impact of metal mixtures is implicated in potentially impairing children's physical development, particularly in terms of HAZ.

Research progress on ion channels and their molecular regulatory mechanisms in the human sperm flagellum.

The ion channels in sperm tail play an important role in triggering key physiological reactions, e.g., progressive motility, hyperactivation, required for successful fertilization. Among them, CatSper and KSper have been shown to be important ion channels for the transport of Ca2+ and K+ . Moreover, the voltage-gated proton channel Hv1, the sperm-specific sodium-hydrogen exchanger (sNHE), the epithelial sodium channel (ENaC), members of the temperature-sensitive TRP channel family, and the cystic fibrosis transmembrane regulator (CFTR) are also found in the flagellum. This review focuses on the latest advances in ion channels located at the flagellum, describes how they affect sperm physiological function, and summarizes some primary mutual regulation mechanism between ion channels, including PH, membrane potential, and cAMP. These ion channels may be promising targets for clinical application in infertility.

A smart pathogen detector engineered from intracellular hydrogelation of DNA-decorated macrophages.

Bacterial infection is a major threat to global public health, which urgently requires useful tools to rapidly analyze pathogens in the early stages of infection. Herein, we develop a smart macrophage (Mø)-based bacteria detector, which can recognize, capture, enrich and detect different bacteria and their secreted exotoxins. We transform the fragile native Møs into robust gelated cell particles (GMøs) using photo-activated crosslinking chemistry, which retains membrane integrity and recognition capacity for different microbes. Meanwhile, these GMøs equipped with magnetic nanoparticles and DNA sensing elements can not only respond to an external magnet for facile bacteria collection, but allow the detection of multiple types of bacteria in a single assay. Additionally, we design a propidium iodide-based staining assay to rapidly detect pathogen-associated exotoxins at ultralow concentrations. Overall, these nanoengineered cell particles have broad applicability in the analysis of bacteria, and could potentially be used for the management and diagnosis of infectious diseases.

Exploring structural and functional alterations in drug-naïve obsessive-compulsive disorder patients: An ultrahigh field multimodal MRI study.

BACKGROUND: Brain structural and functional alterations have been reported in obsessive-compulsive disorder (OCD) patients; however, these findings were inconsistent across studies due to several limitations, including small sample sizes, different inclusion/exclusion criteria, varied demographic characteristics and symptom dimensions, comorbidity, and medication status. Prominent and replicable neuroimaging biomarkers remain to be discovered. METHODS: This study explored the gray matter structure, neural activity, and white matter microstructure differences in 40 drug-naïve OCD patients and 57 matched healthy controls using ultrahigh field 7.0 T multimodal magnetic resonance imaging, which increased the spatial resolution and detection power. We also evaluated correlations among different modalities, imaging features and clinical symptoms. RESULTS: Drug-naïve OCD patients exhibited significantly increased gray matter volume in the frontal cortex, especially in the orbitofrontal cortex, as well as volumetric reduction in the temporal lobe, occipital lobe and cerebellum. Increased neural activities were observed in the cingulate gyri and precuneus. Increased temporal-middle cingulate and posterior cingulate-precuneus functional connectivities and decreased frontal-middle cingulate connectivity were further detected. Decreased fractional anisotropy values were found in the cingulum-hippocampus gyrus and inferior fronto-occipital fascicle in OCD patients. Moreover, significantly altered imaging features were related to OCD symptom severity. Altered functional and structural neural connectivity might influence compulsive and obsessive features, respectively. CONCLUSIONS: Altered structure and function of the classical cortico-striato-thalamo-cortical circuit, limbic system, default mode network, visual, language and sensorimotor networks play important roles in the neurophysiology of OCD.

Amplification-Free, Single-Microbead-Based Cas12a Assay for One-Step DNA Detection at the Single-Molecule Level.

CRISPR/Cas-based systems are highly attractive for developing next-generation diagnostic technologies because of their intrinsic merits such as simplicity, sensitivity, and specificity. However, currently, nucleic acid amplification procedures are still needed to achieve attomolar sensitivity in most CRISPR/Cas-based assays, which causes high cost, operation difficulty, and low efficiency. Herein, we combine the CRISPR/Cas12a-based assay and a single-microbead detection platform for one-step and amplification-free detection of DNA at the single-molecule level. By modifying DNA reporters on a biomimetic membrane-coated microbead, the activated Cas12a by targets will cleave these reporters and lighten the bead within 10 min. The method allows the detection of the target down to three copies in a 5 µL sample. Furthermore, we successfully apply this method for the specific identification of viral infection, foodborne bacteria, and DNA mutation in real samples without extra nucleic acid amplification. We believe that this approach offers new insights for developing CRISPR/Cas-based DNA assays in biomedical applications.

Fabrication of a Polyvalent Aptamer Network on an Electrode Surface for Capture and Analysis of Circulating Tumor Cells.

Capture and analysis of circulating tumor cells (CTCs) from complex matrixes is pivotal for the prediction of cancer metastasis and personalized treatment of cancer. Herein, we propose a strategy for CTC capture by design and fabrication of a polyvalent aptamer network on an electrode surface, which can be further used for the sensitive analysis of CTCs. In our design, the polyvalent aptamer network, which is constructed via a rolling circle amplification reaction, can significantly enhance the cell-binding abilities. Meanwhile, tetrahedral DNA structures previously assembled on the electrode surface will promote the spatial orientation and reduce the steric hindrance effect of the cell capture, thus improving the cell capture efficiency. Importantly, a detectable electrochemical signal can be obtained without additional signal probes by means of target-induced allostery of the DNA hairpin structures. Further studies reveal that the electrochemical response is proportional to the logarithm of the CTC abundance ranging from 102 to 5 × 104 cell mL-1 with a low limit of detection of 23 cell mL-1. Moreover, the proposed capture strategy exhibits excellent stability and anti-interference in human whole blood, indicating its promising potential in clinical diagnosis.

Single-Cell Quantitative Phenotyping via the Aptamer-Mounted Nest-PCR (Apt-nPCR).

Analyzing single-cell phenotypes is increasingly required in biomedical studies, for non-genetic understanding of cellular activities and the biological significance of rare cell subpopulations. However, as compared to the genotypic analysis, single-cell phenotype analysis is technically more challenging. Herein, a tractable method that allows quantitative phenotyping of single cell is developed in this work, termed as the aptamer-mounted nest-PCR (Apt-nPCR). In specific, only two rounds of PCR reactions are required to complete the analysis, where aptamers (short oligonucleotides that bind to specific target molecules) are used as the recognition elements to bind antigens and also as the templates of nPCR for multiplexed and quantitative detection. So, quantitative information of these target antigens can be revealed by quantitative PCR analysis of these aptamers, which can thus be used to interpret cell phenotypes in a quantitative-to-qualitative way. By addressing two technical issues that are involved in single-cell phenotype analysis─multiplexed detection plus high sensitivity, we have shown the availability of this method for single-cell phenotyping. Therefore, the Apt-nPCR method may represent a tractable method to facilitate the single-cell phenotype analysis, which can be used as a complementary method against these single-cell genotyping methods in our daily research.

Demethylation of m1A assisted degradation of the signal probe for rapid electrochemical detection of ALKBH3 activity with practical applications.

ALKBH3 is an important marker for early diagnosis and histopathological grading of prostate cancer. However, the lack of a rapid and sensitive method to quantify the enzyme's activity in the current time necessitates the development of a new quantitative assay. Herein, we first tried to quantitative assay for ALKBH3 activity using an electrochemical method based on the degradation of the signal probe due to alkyl group of the m1A removal by ALKBH3. A strong electrochemical signal can be obtained when the ferrocene (Fc) labeled dsDNAs with 1-methyladenine are immobilized on the electrode. In the presence of ALKBH3, the 3' blunt of DNA can be formed because of the removal of alkyl group of the Fc-DNA probe, which can be recognized and degraded by Exonuclease III (Exo III). As a result, the electrochemical signal produced by Fc greatly decreases, and the activity of ALKBH3 can be easily detected via changes in electrochemical signal. Quantitative analysis of ALKBH3 activity showed a wide detection range (0.1 and 20 ng/mL) and low detection limit (0.04 ng/mL). Furthermore, the method can be applied to detect 1-methyladenine through ALKBH3 in cell lysates and tissue samples, providing a new method for clinical detection of prostate cancer.

Transcriptome profiling of lncRNA and co-expression network in the vaginal epithelial tissue of women with lubrication disorders.

BACKGROUND: Vaginal lubrication is a crucial physiological response that occurs at the beginning of sexual arousal. However, research on lubrication disorders (LD) is still in its infancy, and the role of long non-coding RNAs (lncRNAs) in LD remains unclear. This study aimed to explore the function of lncRNAs in the pathogenesis of vaginal LD. METHODS: The expression profiles of LD and normal control (NC) lncRNAs were examined using next-generation sequencing (NGS), and eight selected differentially expressed lncRNAs were verified by quantitative real-time PCR. We conducted GO annotation and KEGG pathway enrichment analyses to determine the principal functions of significantly deregulated genes. LncRNA-mRNA co-expression and protein-protein interaction (PPI) networks were constructed and the lncRNA transcription factors (TFs) were predicted. RESULTS: From the results, we identified 181,631 lncRNAs and 145,224 mRNAs in vaginal epithelial tissue. Subsequently, our preliminary judgment revealed a total of 499 up-regulated and 337 down-regulated lncRNAs in LD. The top three enriched GO items of the dysregulated lncRNAs included the following significant terms: "contractile fiber part," "actin filament-based process," and "contractile fiber". The most enriched pathways were "cell-extracellular matrix interactions," "muscle contraction," "cell-cell communication," and "cGMP-PKG signaling pathway". Our results also showed that the lncRNA-mRNA co-expression network was a powerful platform for predicting lncRNA functions. We determined the three hub genes, ADCY5, CXCL12, and NMU, using PPI network construction and analysis. A total of 231 TFs were predicted with RHOXF1, SNAI2, ZNF354C and TBX15 were suspected to be involved in the mechanism of LD. CONCLUSION: In this study, we constructed the lncRNA-mRNA co-expression network, predicted the lncRNA TFs, and comprehensively analyzed lncRNA expression profiles in LD, providing a basis for future studies on LD clinical biomarkers and therapeutic targets. Further research is also needed to fully determine lncRNA's role in LD development.

Template-free multiple signal amplification for highly sensitive detection of cancer cell-derived exosomes.

In this work, we have designed a template-free multiple signal amplification method for the highly sensitive detection of cancer cell-derived exosomes. In this design, DNase I serves as a bridge to link the DNA-based amplification approach and terminal deoxynucleotidyl transferase (TdT)-mediated polymerization reaction. Consequently, a detection limit of 10 particles per µL can be achieved, while a complex nucleic acid sequence design can be avoided. This method also exhibits good performance in a complicated matrix and enables the differentiation of healthy individuals from colorectal cancer (CRC) patients.

Does Environmental Policy Affect Green Total Factor Productivity? Quasi-Natural Experiment Based on China's Air Pollution Control and Prevention Action Plan.

It is the scientific way to promote the transformation and optimization of an industrial structure to promote the improvement of its green total factor productivity (GTFP) by formulating environmental regulation policies. Based on the GTFP panel data of 30 provinces in China from 2004 to 2017, this paper takes the "Air Pollution Control and Prevention Action Plan" (APCP Action Plan) as the proxy dummy variable of environmental regulation, and uses the difference-in-differences (DID) model to explore the impact of the implementation of the APCP Action Plan on GTFP. In addition, by constructing the industrial structure optimization index, this paper analyzes how the APCP Action Plan policy affects GTFP through the transformation and optimization of industrial structure. The following basic conclusions are obtained: First, environmental regulation policies like the APCP Action Plan can improve GTFP. Second, the APCP Action Plan has regional heterogeneity in promoting GTFP in different regions. The policy only significantly affects the GTFP in the Pearl River Delta region in southern China. Third, the "quantity" and "quality" of the optimization of industrial structure will weaken the promoting effect of the APCP Action Plan on GTFP. In contrast, the rationalization of industrial structure will aggravate this promoting effect.

Overexpressing miR-122-5p Inhibits the Relaxation of Vaginal Smooth Muscle in Female Sexual Arousal Disorder by Targeting Vasoactive Intestinal Peptide Receptor 1.

INTRODUCTION: Female sexual arousal disorder (FSAD) is a common issue causing physical and psychological pain, but it has no standard diagnostic criteria or treatment. So its pathogenesis desiderates to be explored. AIM: To investigate the specific function of miR-122-5p in FSAD. METHODS: 18 subjects were grouped into FSAD and normal control groups according to the Chinese version of the Female Sexual Function Index, and the expression levels of miR-122-5p and vasoactive intestinal peptide receptor 1 (VIPR1) protein in their tissue were verified through real-time quantitative polymerase chain reaction (qRT-PCR) and western blot (WB) analysis. Then in vitro experiment, miR-122-5p was overexpressed or inhibited in rat vaginal smooth muscle cells (SMCs). The relaxation of rat vaginal SMCs was reflected by the cell morphology, intracellular free cytosolic calcium ion (Ca2+) levels, cell proliferation and apoptosis, together with the cyclic adenosine monophosphate (cAMP) concentration and protein kinase A (PKA) activities. Additionally, the expression levels of relaxation-related proteins, including VIPR1, stimulatory G protein (Gs), adenylate cyclase (AC), and PKA, were detected based on WB analysis. Furthermore, a rescue experiment that simultaneously overexpressed or silenced miR-122-5p and VIPR1 was conducted, and all the indicators were evaluated. MAIN OUTCOMES MEASURE: The expression level of VIPR1 and downstream proteins, cell morphology, cell proliferation and apoptosis, and intracellular free Ca2+ levels were examined. RESULTS: We verified that women with FSAD had higher miR-122-5p and lower VIPR1 protein. Then overexpressing miR-122-5p decreased relaxation of rat vaginal SMCs, which was manifested as a contractile morphology of cells, an increased intracellular free Ca2+ concentration, and lower cAMP concentration and PKA activity. Moreover, by rescue experiments, we inferred that VIPR1 was the target of miR-122-5p and affected the relaxation function of vaginal SMCs. CONCLUSION: miR-122-5p regulates the relaxation of vaginal SMCs in FSAD by targeting VIPR1, ulteriorly providing an underlying diagnostic and therapeutic target for FSAD. Cong S, Gui T, Shi Q, et al. Overexpressing miR-122-5p Inhibits the Relaxation of Vaginal Smooth Muscle in Female Sexual Arousal Disorder by Targeting Vasoactive Intestinal Peptide Receptor 1. Sex Med 2021;9:100390.

miR-195-5p Regulates the Phenotype Switch of CCSM Cells by Targeting Smad7.

INTRODUCTION: Phenotype switch refers to the process in which smooth muscle cells change from contractile type to synthetic type and acquire the ability of proliferation. Phenotypic transformation involves many changes of cell function, such as collagen deposition and fibrosis, which affect the normal erectile function of penis. AIM: To investigate the role of miR-195-5p in regulating the Phenotype switch of the corpus cavernosum smooth muscle (CCSM) cells. METHODS: A small mother against decapentaplegic 7(Smad7) virus vector and a miR-195-5p mimics or an si-Smad7 viral vector and a miR-195-5p inhibitor were transfected into CCSM cells. The cells were obtained by primary culture of rat corpus cavernosum smooth muscle tissue. Real-time polymerase chain reaction (PCR) experiments, Western blotting, hematoxylin-eosin (HE) staining, transwell experiments, MTT assays, and flow cytometry were used to detect miR-195-5p, Smad7, phenotype switch markers of CCSM cells and related protein expression, as well as changes in cell morphology, migration, proliferation and apoptosis. MAIN OUTCOME MEASURE: To study the regulation of miR-195-5p in CCSM cells by overexpression and silencing strategies. RESULTS: Overexpressed miR-195-5p promoted the transformation of CCSM cells from a contractile type to a synthetic type. Meanwhile, the migration ability and proliferation ability of CCSM cells increased, and the apoptosis rate decreased. The expression-silencing of miR-195-5p gave rise to the opposite effect. The results of the rescue experiment demonstrated that overexpressed Smad7 rescued the inhibitory of the switch of the CCSM cell phenotype from the contractile type to the synthesis type caused by overexpression of miR-195-5p alone. Moreover, the enhancement effect of the migration ability and proliferation ability of CCSM cells was also eliminated, and the apoptosis rate was increased. Silencing miR-195-5p and Smad7 at the same time resulted in the opposite effect. CONCLUSION: miR-195-5p may regulate the phenotype switch of CCSM cells by targeting Smad7. Zhang J, Zhang X, Zhang J, et al. miR-195-5p Regulates the Phenotype Switch of CCSM Cells by Targeting Smad7. Sex Med 2021;9:100349.

Construction of circRNA-miRNA-mRNA Network for Exploring Underlying Mechanisms of Lubrication Disorder.

Lubrication disorder is a common health issue that manifests as insufficient sexual arousal at the beginning of sex. It often causes physical and psychological distress. However, there are few studies on lubrication disorder, and the complexity of circular RNA (circRNA) and the related competing endogenous RNA (ceRNA) network in lubrication disorder is still poorly known. Therefore, this study aims to build a regulatory circRNA-micro (mi)RNA-mRNA network and explore potential molecular markers of lubrication disorder. In the study, 12 subjects were recruited, including 6 in the lubrication disorder group and 6 in the normal control group. RNA sequencing was exploited to identify the expression profiles of circRNA, miRNA and mRNA between two groups, and then to construct the circRNA-miRNA-mRNA networks. The enrichment analyses of the differentially expressed (DE)-mRNAs were examined via Gene Set Enrichment Analysis (GSEA). Furthermore, the expression level and interactions among circRNA, miRNA, and mRNA were validated using real-time quantitative polymerase chain reaction (RT-qPCR) and dual-luciferase reporter assays. In the results, 73 circRNAs, 287 miRNAs, and 354 target mRNAs were differentially expressed between two groups when taking | Log2 (fold change)| > 1 and P-value < 0.05 as criteria, and then the results of GSEA revealed that DE-mRNAs were linked with "vascular smooth muscle contraction," "aldosterone regulated sodium reabsorption," "calcium signaling pathway," etc. 19 target relationships among 5 circRNAs, 4 miRNAs, and 7 mRNAs were found and constructed the ceRNA network. Among them, hsa-miR-212-5p and hsa-miR-874-3p were demonstrated to be related to the occurrence of lubrication disorder. Eventually, consistent with sequencing, RT-qPCR showed that hsa_circ_0026782 and ASB2 were upregulated while hsa-miR-874-3p was downregulated, and dual-luciferase reporter assays confirmed the interactions among them. In summary, the findings indicate that the circRNA-miRNA-mRNA regulatory network is presented in lubrication disorder, and ulteriorly provide a deeper understanding of the specific regulatory mechanism of lubrication disorder from the perspective of the circRNA-miRNA-mRNA network.

Zr4+-mediated hybrid chain reaction and its application for highly sensitive electrochemical detection of protein kinase A.

An electrochemical platform has been developed to detect protein kinase activity through the combined actions of Zr4+ mediated signal transition and hybridization chain reaction (HCR)-stimulated DNAzymes nanowires. First of all, protein kinase A (PKA) phosphorylates substrate peptides immobilized on gold electrode surface. Thereafter, the DNA1 containing 5'-phosphoryl ends is linked to the phosphorylated substrate peptide via the robust phosphate-Zr4+-phosphate linkages. By the introduction of molecular beacons (MBs), the DNA1 can open the hairpin structures of MBs through toehold mediated strand displacement (TMSDR), leading to an autonomous stem-opening process and subsequent assembly of G-quadruplex-containing DNA chains by HCR. After the addition of hemin, the formed HRP-mimicking DNAzymes can catalyze the hydroquinone-H2O2 system to generate amplified electrochemical signals. As expected, this method can achieve ultrahigh analytical performance with a low detection limit of 0.02U/mL and exhibit high cost-savings potential without the need for antibody, protease and labeling. Therefore, this method can serve as a new tool for the assay of protein kinase A and its inhibitor screening in the future.

Enzyme-free electrochemical biosensor based on double signal amplification strategy for the ultra-sensitive detection of exosomal microRNAs in biological samples.

Growing evidence suggests that exosomes-encapsulated miRNAs detection is of paramount significance in early diagnostics of cancer due to protection from degradation by ribonuclease. However, exosomal microRNAs with low abundance and subtle variation have restricted their clinical application. Herein, an electrochemical biosensor for highly sensitive detection of exosomal microRNAs has been fabricated based on the double signal amplification strategy. Our proposed amplifier consists of two steps: target miRNA cyclic signal amplification induced by strand displacement reaction (SDR) and subsequent deposition of silver nanoparticles induced by streptavidin-biotin interaction. Consequently, this method shows ultrahigh sensitivity to detect miRNA. Taking miRNA-21 in exosomes as a model analyte, a detection limit of 0.4 fM (S/N = 3) can be obtained. Meanwhile, the method is relatively simple and low-cost without the requirement of enzyme, which has been applied in biological samples successfully. Therefore, our miRNA assay method has shown great promise as molecular tool in the detection of exosomal miRNA and could be widely used in clinic in the future.

[Differential expression profiles of miRNAs in the vaginal tissue of the women with lubrication disorders].

OBJECTIVE: To explore the pathogenesis of lubrication disorder (LD), a most common type of female sexual dysfunction affecting women's physical health and conception, and find the therapeutic targets for its treatment and prevention. METHODS: We chose 3 LD patients and 3 healthy controls in Nanjing Maternal and Child Health Hospital, extracted their vaginal epithelial RNA for high-throughput miRNA sequencing, screened differentially expressed miRNAs for hierarchical cluster analysis, target gene prediction and gene ontology (GO) and KEGG pathway enrichment analyses. Finally, we verified the sequencing results by real-time fluorescent quantitative PCR. RESULTS: Totally 1 673 miRNAs were predicted by high-throughput sequencing and 64 likely to be the targets for the treatment of LD were screened, including 25 up-regulated more than 4 times and 39 down-regulated more than 4 times in the LD patients compared with the healthy controls. The neuron projection morphogenesis and AMPK signaling pathway were the most significant enrichment GO term and KEGG pathway. CONCLUSIONS: miRNAs are expressed differentially in LD patients. These miRNAs and target genes may be related to the occurrence of LD, and those that are expected to be the targets for the treatment of LD have important theoretical significance and potential application value.

Bio-inspired construction of a semi-artificial enzyme complex for detecting histone acetyltransferases activity.

Herein, an electrochemical method to detect histone acetyltransferases activity (HAT) has been developed based on the reduction of G-Quadruplex-Cu(ii) metalloenzyme activity. A G-quadruplex-Cu(ii) metalloenzyme has excellent peroxidase property, generating strong electrochemical signal. In the presence of HAT, it can catalyze substrate peptide acetylation and produce large amounts of Coenzyme A (CoA). The electrochemical signal of G-Quadruplex-Cu(ii) is weak due to the competitive combination between G-Quadruplex and CoA with Cu(ii), resulting in the direct quantitative detection of HAT. The detection limit for HAT is about 0.14 nM using this strategy and the cost is quite low since the developed assay method is label-free and antibody-free due to the use of low-cost DNA and Cu2+. Since this assay method can be employed to detect HAT in serum, it may be useful in disease diagnosis in the future.