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Single-level biomarker detection has the limitation of insufficient accuracy in cancer diagnosis. Therefore, the strategy of developing highly sensitive, multi-channel biosensors for high-throughput ctDNA determination is critical to improve the accuracy of early diagnosis of clinical tumors. Herein, in order to achieve efficient detection of up to ten targets for early diagnosis of ovarian cancer, a DNA-nanoswitch-based multi-channel (DNA-NSMC) biosensor was built based on the multi-module catalytic hairpin assembly-mediated signal amplification (CHA) and toehold-mediated DNA strand displacement (TDSD) reaction. Only two different fluorescence signals were used as outputs, combined with modular segmentation strategy of DNA-nanoswitch-based reaction platform; the multi-channel detection of up to ten targets was successfully achieved for the first time. The experimental results suggest that the proposed biosensor is a promising tool for simultaneously detecting multiple biomarkers for the early diagnosis of ovarian cancer, offering new strategies for the early screening, diagnosis, and treatment not only for ovarian cancer but also for other cancers., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: Although brain glymphatic dysfunction is a contributing factor to the cognitive deficits in Parkinson's disease (PD), its role in the longitudinal progression of cognitive dysfunction remains unknown., Objective: To investigate the glymphatic function in PD with mild cognitive impairment (MCI) that progresses to dementia (PDD) and to determine its predictive value in identifying individuals at high risk for developing dementia., Methods: We included 64 patients with PD meeting criteria for MCI and categorized them as either progressed to PDD (converters) (n = 29) or did not progress to PDD (nonconverters) (n = 35), depending on whether they developed dementia during follow-up. Meanwhile, 35 age- and gender-matched healthy controls (HC) were included. Bilateral diffusion-tensor imaging analysis along the perivascular space (DTI-ALPS) indices and enlarged perivascular spaces (EPVS) volume fraction in bilateral centrum semiovale, basal ganglia (BG), and midbrain were compared among the three groups. Correlations among the DTI-ALPS index and EPVS, as well as cognitive performance were analyzed. Additionally, we investigated the mediation effect of EPVS on DTI-ALPS and cognitive function., Results: PDD converters had lower cognitive composites scores in the executive domains than did nonconverters (P < 0.001). Besides, PDD converters had a significantly lower DTI-ALPS index in the left hemisphere (P < 0.001) and a larger volume fraction of BG-PVS (P = 0.03) compared to HC and PDD nonconverters. Lower DTI-ALPS index and increased BG-PVS volume fraction were associated with worse performance in the global cognitive performance and executive function. However, there was no significant mediating effect. Receiver operating characteristic analysis revealed that the DTI-ALPS could effectively identify PDD converters with an area under the curve (AUC) of 0.850., Conclusion: The reduction of glymphatic activity, measured by the DTI-ALPS, could potentially be used as a non-invasive indicator in forecasting high risk of dementia conversion before the onset of dementia in PD patients., (© 2024. The Author(s).)

An electrochemical biosensor has been developed for detection of Escherichia coli O157 by integrating lateral flow with screen-printed electrodes. The screen-printed electrodes were attached under the lateral flow detection line, and organic-inorganic nanoflowers prepared from E. coli O157-specific antibodies as an organic component were attached to the lateral flow detection line. In the presence of E. coli O157, an organic-inorganic nanoflower-E. coli O157-antimicrobial peptide-labelled ferrocene sandwich structure is formed on the lateral flow detection line. Differential pulse voltammetry is applied using a smartphone-based device to monitor ferrocene on the detection line. The resulting electrochemical biosensor could specifically detect E. coli O157 with a limit of detection of 25 colony-forming units mL -1 . Through substitution of antibodies of organic components in organic-inorganic nanoflowers, biosensors have great potential for the detection of other pathogens in biomedical research and clinical diagnosis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Single nucleotide polymorphism (SNP) biosensors are emerging rapidly for their promising applications in human disease prevention diagnosis, treatment, and prognosis. However, it remains a bottleneck in equipping simple and stable biosensors with the traits of high sensitivity, non-enzyme, and low cost. Double base mismatches mediated chain displacement reactions have attracted fascinating advantages of tailorable thermodynamics stability, non-enzyme, and excellent assembly compliance to involvement in SNP identification. As the base mismatch position and amount in DNA sequence can be artificially adjusted, it provides plenty of selectivity and specificity for exploring perfect biosensors. Herein, a biosensor with double base mismatches mediated catalytic hairpin assembly (CHA) is designed via one base mismatch in the toehold domain and the other base mismatch in the stem sequence of hairpin 1 (H1) by triggering CHA reaction to achieve selective amplification of the mutation target (MT) and fluorescence resonance energy transfer (FRET) effect that is composed of Cy3 and Cy5 terminally attached H1 and hairpin 2 (H2). Depending on the rationally designed base mismatch position and toehold length, the fabricated biosensors show superior SNP detection performance, exhibiting a good linearity with high sensitivity of 6.6 fM detection limit and a broad detection abundance of 1%. The proposed biosensor can be used to detect the KRAS mutation gene in real samples and obtain good recoveries between 106 and 116.99%. Remarkably, these extendible designs of base mismatches can be used for more types of SNP detection, providing flexible adjustment based on base mismatch position and toehold length variations, especially for their thermodynamic model for DNA-strand displacement reactions., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

We propose a sensitive H1N1 virus fluorescence biosensor based on ligation-transcription and CRISPR/Cas13a-assisted cascade amplification strategies. Products are generated via the hybridization of single-stranded DNA (ssDNA) probes containing T7 promoter and crRNA templates to a target RNA sequence using SplintR ligase. This generates large crRNA quantities in the presence of T7 RNA polymerase. At such crRNA quantities, ternary Cas13a, crRNA, and activator complexes are successfully constructed and activate Cas13a to enhance fluorescence signal outputs. The biosensor sensitively and specifically monitored H1N1 viral RNA levels down to 3.23 pM and showed good linearity when H1N1 RNA concentrations were 100 pM-1 µM. Biosensor specificity was also excellent. Importantly, our biosensor may be used to detect other viral RNAs by altering the sequences of the two probe junctions, with potential applications for the clinical diagnosis of viruses and other biomedical studies., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Purpose: Dysfunctions of retinal pigment epithelium (RPE) attributed to oxidative stress and inflammation are implicated with age-related macular degeneration (AMD). A debate on the curative role of metformin in AMD has been raised, though several recent clinical studies support the lower odds by using metformin. This study aimed to determine whether metformin could exert cytoprotection against RPE oxidative damages and the potential mechanisms., Methods: A cellular AMD model was established by treating ARPE-19 cells with hydrogen peroxide (H 2 O 2 ) for 24 h. The reactive oxygen species (ROS) generation, expression of antioxidant enzymes, and levels of pro-inflammatory cytokines were monitored under administrations with H 2 O 2 with/without metformin. The expression and DNA-binding activity of transcription factor erythroid-related factor 2 (Nrf2) were determined by western blot, immunofluorescence, and electrophoretic mobility shift assay. Knockout of Nrf2 was conducted by CRISPR/Cas9 gene deletion system., Results: Metformin pretreatment significantly improved the H 2 O 2 -induced low viability of ARPE-19 cells, reduced ROS production, and increased contents of antioxidative molecules. Concurrently, metformin also suppressed levels of pro-inflammatory cytokines caused by H 2 O 2 . The metformin-augmented nuclear translocation and DNA-binding activity of Nrf2 were further verified by the increased expression of its downstream targets. Genetic deletion of Nrf2 blocked the cytoprotective role of metformin., Conclusion: Metformin possesses antioxidative and anti-inflammatory properties in ARPE-19 cells by activating the Nrf2 signaling. It supports the potential use for the control and prevention of AMD., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study provides proof of concept of a colorimetric biosensor for influenza H1N1 virus assay based on the CRISPR/Cas13a system and hybridization chain reaction (HCR). Target RNA of influenza H1N1 virus activated the trans-cleavage activity of Cas13a, which cleaved the special RNA sequence (-UUU-) of the probe, further initiating HCR to copiously generate G-rich DNA. Abundant G-quadruplex/hemin was formed in the presence of hemin, thus catalyzing a colorimetric reaction. The colorimetric biosensor exhibited a linear relationship from 10 pM to 100 nM. The detection limit was 0.152 pM. The biosensor specificity was excellent. This new and sensitive detection method for influenza virus is a promising rapid influenza diagnostic test., (© 2022. Springer-Verlag GmbH Germany, part of Springer Nature.)

Dihydro-β-ionone is a characteristic aroma compound of Osmanthus fragrans and is widely applied in the flavor & fragrance industry. However, the main focus is on chemical synthesis due to the metabolic pathways of dihydro-β-ionone is still unclear. Here, we explored the one-pot synthesis system for dihydro-β-ionone production using carotenoid cleavage dioxygenase (CCD) and enoate reductase. After screening the CCD enzyme, PhCCD1 from the Petunia hybrid was identified as the suitable enzyme for the first step of dihydro-β-ionone synthesis due to the high enzyme activity for carotenoid. The PhCCD1 was expressed in Escherichia coli and further characterized. The optimal activity of PhCCD1 was observed at pH 6.8 and 45 °C. The enzyme was stable over the pH range of 6.0-8.0 and had good thermal stability below 40 °C. Then, we optimized the coupled reaction conditions for dihydro-β-ionone production by PhCCD1 and enoate reductase AaDBR1 from Artemisia annua. Furthermore, we introduced the NADPH regeneration system with a 1.5-fold enhancement for dihydro-β-ionone production. Collectively, approximately 13.34 mg/L dihydro-β-ionone was obtained by the one-pot biosystem with a corresponding molar conversion of 85.8%. For the first time, we successfully designed and constructed a new synthesis pathway for dihydro-β-ionone production in vitro. The coupled catalysis reported herein illustrates the feasibility of producing dihydro-β-ionone from carotenoids and guides further engineering in the food industry., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In this study, Lba Cas12a (Cpf1) as one of the CRISPR systems from Lachnospiraceae bacterium was coupled with a hybridization chain reaction (HCR) to develop an electrochemical biosensor for detecting the pathogenic bacterium, Salmonella typhimurium. Autonomous cross-opening of functional DNA hairpin structures of HCR yielded polymer double-stranded DNA wires consisting of numerous single-stranded DNAs, which initiated the trans-cleavage activity of CRISPR-Cas12a to indiscriminately cleave random single-stranded DNA labeling electrochemical tags on the surface of the electrode. It led to a variation in the electron transfer of electrochemical tags. The polymer double-stranded DNA of HCR was immobilized on dynabeads (DBs) via the S. typhimurium aptamer and released from DBs. The established method could selectively and sensitively quantify S. typhimurium in samples with detection limits of 20 CFU/mL. Our study provides a novel insight for exploring universal analytical methods for pathogenic bacteria based on CRISPR-Cas12a coupled with HCR., (© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

Understanding dynamic changes in the genetic architecture of quantitative traits is crucial in developmental genetics. Functional mapping is an appropriate method that passes a mathematical equation to describe a biological developmental process with the genetic mapping framework. Appropriate genetic model and applicable mapping population are indispensable condition for functional mapping of important agronomic traits in plants. Based on the Wang-Lan-Ding model, we ever applied a DH population to carry out functional mapping QTLs underlying growth trajectory on tiller number. However, inconsistent genetic background among the DH lines might disturb the mapping results. With the advent of innovative research materials, single segment substitution lines, allows us to do more precise genetic analyses. Thus functional mapping was again conducted on tiller number using the Wang-Lan-Ding model and a single segment substitution line population with the genetic background of Huajingxian 74 so as to explore QTL dynamic mechanism to regulate developmental traits. We detected that all five single segment substitution lines harbored tillering QTLs with additives and/or dominances to influence the four functional parameters, the optimum tillering time (t 0 ), the maximum tiller number (K), the tillering increased rate (r) and the tillering decreased rate (c), which were estimated from the Wang-Lan-Ding model and with some biological meaning. They mainly brought the inflexion point (t 0 ) delay, the peak increase (K) and the degradation (c) acceleration, while the growth (r) slow down. Moreover, epistatic interactions among these QTLs were confirmed to be prevalent. A total of 39 significant epistatic effects were detected to associate with the four parameters, occupying 34.8% of 112 pairs of epistatic interactions investigated. Contrary to the QTL effects, these epistatic effects largely decreased t 0 , K and c, while increased r. Our results indicated that the five QTL effects and their epistatic effects significantly changed the shape and trajectory of tiller number via influence of the four functional parameters. Rational use of these QTLs is expected to improve tillering number of rice by molecular design breeding., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A point-of-care (POC) immunoassay was established for the sensitive and rapid detection of pathogenic Escherichia coli O157:H7, using magnetic Fe 3 O 4 organic-inorganic composites (Ab@Fe 3 O 4 ) for immunomagnetic separation, nanozyme platinum nanoparticle (PtNp) organic-inorganic composites (Ap@PtNp) for signal amplification, and thermometer readings. Antibodies and Fe 3 O 4 were incubated in Cu 2+ phosphate buffer to synthesize the magnetic composite Ab@Fe 3 O 4 with antibodies, to specifically capture E. coli O157:H7. Antimicrobial peptides and PtNp were incubated in Cu 2+ phosphate buffer to synthesize the signal composites Ap@PtNp with antimicrobial peptides (magainin I), recognizing and labeling E. coli O157:H7. In the presence of E. coli O157:H7, magnetic microcomposites targeted bacteria and signal microcomposites to form the sandwich structure: Ab@Fe 3 O 4 -bacteria-Ap@PtNp for magnetic separation. Ap@PtNp of signal composites catalyzed H 2 O 2 to generate thermo-signals (temperature rise), which were determined by a thermometer. This point-of-care bioassay detected E. coli O157:H7 in the linear range of 10 1 -10 7  CFU mL -1 and with a detection limit of 14 CFU mL -1 . One-pot process magnetic Fe 3 O 4 organic-inorganic composites (Ab@Fe 3 O 4 , magnetic microcomposites, MMC) for immunomagnetic separation and nanozyme platinum nanoparticle (PtNp) organic-inorganic composites (Ap@PtNp, signal microcomposites, SMC) were used as signal amplification and thermometer readings for E. coli O157:H7 detection.

Aldehyde dehydrogenase (ALDH) was first developed as an enzymatic signaling system of a biosensor for sensitive point-of-care detection of pathogenic bacteria. ALDH and specific aptamers to Salmonella typhimurium (S. typhimurium), as organic components, were embedded in organic-inorganic nanocomposites as a biosensor signal label, integrating the functions of signal amplification and target recognition. The biosensing mechanism is based on the fact that ALDH can catalyze rapid oxidation of acetaldehyde into acetic acid, resulting in pH change with portable pH meter readout. The altered pH exhibited a linear relationship with the logarithm of S. typhimurium from 10 2 to 10 8  CFU/mL and detection limit of 46 CFU/mL. Thus, the proposed biosensor has potential application in the diagnosis of pathogenic bacteria.

A sandwich immunoassay was developed for determination of E. coli O157:H7. This is based on an antimicrobial peptide-mediated nanocomposite pair and uses a personal glucose meter as signal readout. The antimicrobial peptides, magainins I, and cecropin P1 were employed as recognition molecules for the nanocomposite pair, respectively. With a one-step process, copper phosphate nanocomposites embedded by magainins I and Fe 3 O 4 were used as "capturing" probes for bacterial magnetic isolation, and calcium phosphate nanocomplexes composed of cecropin P1 and invertase were used as signal tags. After magnetic separation, the invertase of the signal tags hydrolyzed sucrose to glucose, thereby converting E. coli O157:H7 levels to glucose levels. This latter can be quantified by a personal glucose meter. Under optimal conditions, the concentration of E. coli O157:H7 can be determined in a linear range of 10 to 10 7  CFU·mL -1 with a detection limit of 10 CFU·mL -1 . The method was successfully applied to the determination of E. coli O157:H7 in milk samples. Graphical abstract Schematic representation of sandwich immunoassay for E. coli O157:H7. One-pot synthetic of Fe 3 O 4 -magainins I nanocomposites (MMP) were used for magnetic capture. Cecropin P1-invertase nanocomposites (PIP) were used as signal tags. A personal glucose meter was used as readout to determine the target.

The published version of this article, unfortunately, contained error. The authors are re-writing to express their sincere apology for a mistake that a mark "10 -5 , 10 -4 , 10 -3 , 10 -2 , 10 -1 CFU•mL -1 " in the legend of Fig. 2 was not corrected as "10 5 , 10 4 , 10 3 , 10 2 , 10 1 CFU•mL -1 ".

Pregnancy test strips are widely used in daily life. A commercial pregnancy test strip was modified to obtain a point-of-care device for the detection of pathogenic bacteria. Hybrid nanoflowers were prepared from concanavalin A, human chorionic gonadotropin, and Cu 3 (PO 4 ) 2 via a one-pot method. They were used as signaling probes in an off-the-shelf pregnancy test strip. This modified lateral flow immunoassay can detect Escherichia coli O157:H7 with a detection limit of 4 CFU·mL -1 , and Salmonella typhimurium with a detection limit of 3 CFU·mL -1 . Conceivably, the method has high potential as a portable and cost-effective tool for rapid determination of a wide range of analytes, especially in resource-constrained settings. Graphical abstract Hybrid nanoflower loaded human chorionic gonadotropin (hCG) and concanavalin A (hCG - nanoflowers) were synthesized via a one-pot method and used as signal labels with commercial commercial-off-the-shelf pregnancy test strips to detect pathogenic bacteria targets, thus yielding an easily smartphone readout signal.

Purpose: Pelvic lymph node dissection (PLND) has been performed during radical prostatectomy in nearly all patients with clinically localized prostatic carcinoma (PCa), while the specific regions that needed to be removed demonstrated bifurcation among urologist. However, clinical studies comparing extended PLND (ePLND) with standard PLND (sPLND) and limited PLND (lPLND) reveal conflicting, or even opposing results., Methods: All controlled trials comparing ePLND with sPLND or lPLND were identified through comprehensive searches of the PubMed, Cochrane Library and Embase databases. A systematic review and meta-analysis of these studies were then performed., Results: Eighteen studies with a total of 8,914 patients were included. Regardless of being compared with sPLND or lPLND, ePLND significantly improved LN retrieval [ePLND vs. sPLND: weighted mean difference (WMD) 11.93, 95 % confidence interval (CI) 9.91-13.95, p < 0.00001; ePLND vs. lPLND: WMD 8.27, 95 % CI 3.53-13.01, p = 0.0006] and the detection of more LNs positive of metastasis [risk ratio (RR) 3.51, 95 % CI 2.14-5.75, p < 0.00001; RR 3.50, 95 % CI 2.20-5.55, p < 0.00001, respectively]. EPLND decreased the complication rate, but the differences were not statistically significant (RR 1.52, 95 % CI 0.87-2.65, p = 0.14; RR 1.52, 95 % CI 0.67-3.45, p = 0.32, respectively). Operating time, estimated blood loss, length of hospital stay and biochemical recurrence (BCR) were statistically insignificant between techniques., Conclusions: ePLND shows benefits associated with increased LNs yield, LNs positivity, and safety, significantly with no risk of side effects. However, ePLND did not decrease BCR. Additional high-quality, well-designed randomized controlled trials and comparative studies with long-term follow-up results are required to define the optimal procedure for patients with clinically localized PCa.

Postmenopausal osteoporosis is associated with high level of adipogenesis within the bone marrow at the expense of osteoblast population. The mechanical effect on β-catenin through phosphorylation of glycogen synthase kinase-3β (GSK-3β) is critical for inhibition of adipogenesis in mesenchymal stem cells in vitro. In present study, we hypothesized that treadmill training could regulate the β-catenin signaling through phosphorylation of GSK-3β in the lumbar vertebrae of ovariectomized (OVX) rats. 3-month-old female Sprague-Dawley rats were divided randomly into the following four groups: (a) Sham, (b) OVX, (c) OVX exercised (EX), and (d) OVX estrogen replacement (E(2)). At the end of the experiment, the serum levels of estradiol (E(2)) and luteinizing hormone (LH), the ultimate lumbar vertebra strength, as well as the protein expression for peroxisome proliferators-activated receptor γ (PPARγ), β-catenin, P-GSK-3β, and osterix (Osx) in lumbar vertebrae were analyzed. Moreover, the protein expression for β-catenin and P-GSK-3β were also examined in the uterus. The EX group had lower protein level of PPARγ, higher ultimate lumbar vertebral strength, and higher protein levels of β-catenin, and P-GSK-3β in lumbar vertebral bodies compared with sedentary OVX group. The effects of EX treatment on the protein levels of β-catenin and P-GSK-3β in bones were not reproducible in the uterus. Moreover, exercise treatment produced no estrogenic effect as evidenced by serum level of LH. In conclusion, this study suggested that treadmill training could activate the GSK-3β/β-catenin signaling and inhibit the production of PPARγ in lumbar vertebrae of OVX rats, which may contribute to the prevention of bone loss in OVX rats.

Exercise training has been reported to prevent bone loss in ovariectomized (OVX) rats and postmenopausal women. We hypothesized that treadmill training inhibited adipogenesis and enhanced osteogenesis through the regulation of adipocyte differentiation factor peroxisome proliferators-activated receptor gamma (PPARγ) and the osteogenic factor runt-related transcription factor 2 (Runx2) in a model of OVX-induced osteoporosis. To test this hypothesis, 3-month-old female Sprague-Dawley rats were divided randomly into the following groups: Sham, OVX, OVX exercised (EX), and OVX estrogen replacement (E(2)). At the end of the experiment, the bone mineral density (BMD) was detected using DEXA and the morphology change of bone tissues and uterus was observed by HE staining. The protein expression for PPARγ and Runx2 were measured by immunohistochemistry and western blot and the bone triacylglycerol (TG) was extracted by methanol/chloroform. OVX dramatically increased the number of fat vacuoles, protein levels for PPARγ and Runx2 as well as the TG level in tibiae and lumbar vertebrate. In contrast, the serum level of E(2), the lumbar vertebrate BMD as well as the proximal and distal femur BMD was significantly decreased in the OVX group. All changes induced by OVX were significantly reversed by exercise treatment except for the protein expression level of Runx2. Moreover, exercise treatment produced no estrogenic effects on uterus as evidenced by the uterus wet weight and histology. Treadmill training could prevent bone loss induced by OVX through the inhibition of adipocyte differentiation factor PPARγ rather than promoting osteogenic factor Runx2.

To explore the mechanism(s) of exercise training on ovariectomized (OVX)-induced liver lipid disorder, we observed effects of treadmill training on liver fat accumulation and ER alpha expression in intact and ovariectomized rats. Sixty female rats were randomly assigned to six groups: Sham sedentary (S-S), Sham exercised (S-EX), ovariectomized sedentary (O-S), ovariectomized exercised (O-EX), ovariectomized injected subcutaneously with 17beta-estradiol (E(2)) (O-E(2)), and ovariectomized treated with E(2) and exercise (O-E(2)-EX). Twelve weeks after intervention, OVX resulted in significantly higher body weight gain, intra-abdominal fat mass, serum levels of total cholesterol (TC), and liver triacylglycerol (TAG) concentrations and ER alpha expression than S-S group, while the relative uterus and liver mass, serum levels of E(2), TAG, and the ratio of high density lipoprotein (HDL) to TC were markedly lower in O-S group. All of these changes were decreased in O-S rats after treatment with E(2) alone with the exception of serum TC and HDL-C levels and liver ER alpha expression. Exercise alone significantly reversed the effect of OVX on serum E(2), the ratio of HDL-C to TC and the liver and intra-abdominal fat accumulation in OVX rats. The addition of E(2) to exercise induced the same uterus and lipid profile as E(2) alone. Moreover, an additive effect of exercise and E(2) was observed on liver ER alpha expression in Sham or OVX rats. In conclusion, treadmill training alone could prevent liver fat accumulation in OVX rats and the regulation of exercise on liver ER alpha expression in both OVX and Sham rats needs the presence of physical estrogen levels.