Characterization and quantification of silver complexes with dissolved organic matter by size exclusion chromatography coupled to ICP-MS.

The fate and behavior of silver in aquatic systems is intricately determined by its interactions with dissolved organic matter (DOM). In this study, we have introduced a method for identification and quantification of silver-DOM complexes using size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS). Our findings revealed that silver(I) was weakly bound to Suwannee River humic acid, fulvic acid, and natural organic matter (SRHA, SRFA, and SRNOM) in various media, resulting in facile dissociation during chromatographic separation. Suitable chromatographic conditions were determined for the elution of Ag-DOM complexes, involving the use of 0.5 mM ammonium acetate (pH 7) as the mobile phase and silver-aged column (pre-absorbing 0.1-0.7 µg silver(I)). SEC-UV and SEC-ICP-MS chromatograms revealed that Ag-binding fractions of DOM were dominated by its aromatic compounds. The quantification of silver-DOM complexes was achieved by SEC-ICP-MS combination with on-line isotope dilution. Silver at concentrations below 20 µg L-1 was mainly present in the form of organic complexes in low salinity water. These measurements aligned well with the results obtained using the equilibrium dialysis method. Species analyses of Ag-DOM complexes provide a deeper understanding of the reactivity, transport, and fate of silver in aquatic environments. ENVIRONMENTAL IMPLICATION: Ionic silver is highly toxic to aquatic organisms such as fish and zooplankton. The complexation of silver with binding sites within DOM significantly influences its speciation, mobility, and toxicity. Despite the complex and unknown structure of silver-DOM complexes, this study provided a SEC-ICP-MS method to identify and quantify these complexes in a range of media. By uncovering the formation of silver-DOM complexes across diverse media, this work enhances the comprehension of silver transformation processes and associated environmental risks in aquatic environments.

Learning to Guide Particle Search for Dynamic Multiobjective Optimization.

Dynamic multiobjective optimization problems (DMOPs) are characterized by multiple objectives that change over time in varying environments. More specifically, environmental changes can be described as various dynamics. However, it is difficult for existing dynamic multiobjective algorithms (DMOAs) to handle DMOPs due to their inability to learn in different environments to guide the search. Besides, solving DMOPs is typically an online task, requiring low computational cost of a DMOA. To address the above challenges, we propose a particle search guidance network (PSGN), capable of directing individuals' search actions, including learning target selection and acceleration coefficient control. PSGN can learn the actions that should be taken in each environment through rewarding or punishing the network by reinforcement learning. Thus, PSGN is capable of tackling DMOPs of various dynamics. Additionally, we efficiently adjust PSGN hidden nodes and update the output weights in an incremental learning way, enabling PSGN to direct particle search at a low computational cost. We compare the proposed PSGN with seven state-of-the-art algorithms, and the excellent performance of PSGN verifies that it can handle DMOPs of various dynamics in a computationally very efficient way.

Generation of SARS-CoV-2 spike receptor binding domain mutants and functional screening for immune evaders using a novel lentivirus-based system.

The emergence of rapid and continuous mutations of severe acute respiratory syndrome 2 (SARS-CoV-2) spike glycoprotein that increased with the Omicron variant points out the necessity to anticipate such mutations for conceiving specific and adaptable therapies to avoid another pandemic. The crucial target for the antibody treatment and vaccine design is the receptor binding domain (RBD) of the SARS-CoV-2 spike. It is also the site where the virus has shown its high ability to mutate and consequently escape immune response. We developed a robust and simple method for generating a large number of functional SARS-CoV-2 spike RBD mutants by error-prone PCR and a novel nonreplicative lentivirus-based system. We prepared anti-RBD wild type (WT) polyclonal antibodies and used them to screen and select for mutant libraries that escape inhibition of virion entry into recipient cells expressing human angiotensin-converting enzyme 2 and transmembrane serine protease 2. We isolated, cloned, and sequenced six mutants totally bearing nine mutation sites. Eight mutations were found in successive WT variants, including Omicron and other recombinants, whereas one is novel. These results, together with the detailed functional analyses of two mutants provided the proof of concept for our approach.

Development of an Alternative In Vitro Rumen Fermentation Prediction Model.

The aim of this study is to identify an alternative approach for simulating the in vitro fermentation and quantifying the production of rumen methane and rumen acetic acid during the rumen fermentation process with different total mixed rations. In this experiment, dietary nutrient compositions (neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), and dry matter (DM)) were selected as input parameters to establish three prediction models for rumen fermentation parameters (methane and acetic acid): an artificial neural network model, a genetic algorithm-bp model, and a support vector machine model. The research findings show that the three models had similar simulation results that aligned with the measured data trends (R2 ≥ 0.83). Additionally, the root mean square errors (RMSEs) were ≤1.85 mL/g in the rumen methane model and ≤2.248 mmol/L in the rumen acetic acid model. Finally, this study also demonstrates the models' capacity for generalization through an independent verification experiment, as they effectively predicted outcomes even when significant trial factors were manipulated. These results suggest that machine learning-based in vitro rumen models can serve as a valuable tool for quantifying rumen fermentation parameters, guiding the optimization of dietary structures for dairy cows, rapidly screening methane-reducing feed options, and enhancing feeding efficiency.

A nanobody inhibiting porcine reproductive and respiratory syndrome virus replication via blocking self-interaction of viral nucleocapsid protein.

Porcine reproductive and respiratory syndrome (PRRS) is a serious global pig industry disease. Understanding the mechanism of viral replication and developing efficient antiviral strategies are necessary for combating with PRRS virus (PRRSV) infection. Recently, nanobody is considered to be a promising antiviral drug, especially for respiratory viruses. The present study evaluated two nanobodies against PRRSV nucleocapsid (N) protein (PRRSV-N-Nb1 and -Nb2) for their anti-PRRSV activity in vitro and in vivo. The results showed that intracellularly expressed PRRSV-N-Nb1 significantly inhibited PRRSV-2 replication in MARC-145 cells (approximately 100%). Then, the PRRSV-N-Nb1 fused with porcine IgG Fc (Nb1-pFc) as a delivering tag was produced and used to determine its effect on PRRSV-2 replication in porcine alveolar macrophages (PAMs) and pigs. The inhibition rate of Nb1-pFc against PRRSV-2 in PAMs could reach >90%, and it can also inhibit viral replication in vivo. Epitope mapping showed that the motif Serine 105 (S105) in PRRSV-2 N protein was the key amino acid binding to PRRSV-N-Nb1, which is also pivotal for the self-interaction of N protein via binding to Arginine 97. Moreover, viral particles were not successfully rescued when the S105 motif was mutated to Alanine (S105A). Attachment, entry, genome replication, release, docking model analysis, and blocking enzyme-linked immunosorbent assay (ELISA) indicated that the binding of PRRSV-N-Nb1 to N protein could block its self-binding, which prevents the viral replication of PRRSV. PRRSV-N-Nb1 may be a promising drug to counter PRRSV-2 infection. We also provided some new insights into the molecular basis of PRRSV N protein self-binding and assembly of viral particles.IMPORTANCEPorcine reproductive and respiratory syndrome virus (PRRSV) causes serious economic losses to the swine industry worldwide, and there are no highly effective strategies for prevention. Nanobodies are considered a promising novel approach for treating diseases because of their ease of production and low costing. Here, we showed that PRRSV-N-Nb1 against PRRSV-N protein significantly inhibited PRRSV-2 replication in vitro and in vivo. Furthermore, we demonstrated that the motif Serine 105 (S105) in PRRSV-N protein was the key amino acid to interact with PRRSV-N-Nb1 and bond to its motif R97, which is important for the self-binding of N protein. The PRRSV-N-Nb1 could block the self-interaction of N protein following viral assembly. These findings not only provide insights into the molecular basis of PRRSV N protein self-binding as a key factor for viral replication for the first time but also highlight a novel target for the development of anti-PRRSV replication drugs.

MscI restriction enzyme cooperating recombinase-aided isothermal amplification for the ultrasensitive and rapid detection of low-abundance EGFR mutations on microfluidic chip.

The detection of low-abundance mutation genes of the epidermal growth factor receptor (EGFR) exon 21 (EGFR L858R) plays a crucial role in the diagnosis of non-small cell lung cancer (NSCLC), as it enables early cancer detection and facilitates the development of treatment strategies. A detection platform was developed by combining the MscI restriction enzyme with the recombinase-aided isothermal amplification (RAA) technique (MRE-RAA). During the RAA process, "TGG^CCA" site of the wild-type genes was cleaved by the MscI restriction enzyme, while only the low-abundance mutation genes underwent amplification. Notably, when the RAA product was combined with CRISPR-Cas system, the sensitivity of detecting the EGFR L858R mutation increased by up to 1000-fold for addition of the MscI restriction enzyme. This achievement marked the first instance of attaining an analytical sensitivity of 0.001%. Furthermore, a disk-shaped microfluidic chip was developed to automate pretreatment while concurrently analyzing four blood samples. The microfluidic features of the chip include DNA extraction, MRE-RAA, and CRISPR-based detection. The fluorescence signal is employed for detection in the microfluidic chip, which is visible to the naked eye upon exposure to blue light irradiation. Furthermore, this platform has the capability to facilitate early diagnosis for various types of cancer by enabling high-sensitivity detection of low-abundance mutation genes.

Simultaneous removal of calcium, cadmium and tetracycline from reverse osmosis wastewater by sycamore deciduous biochar, shell powder and polyurethane sponge combined with biofilm reactor.

The treatment of reverse osmosis concentrate generated from urban industrial sewage for resource recovery has been hot. In this research, a biofilm reactor was constructed by combining sycamore deciduous biochar, shell powder, and polyurethane sponge loaded with Zobellella denitrificans sp. LX16. For ammonia nitrogen (NH4+-N), calcium (Ca2+), chemical oxygen demand (COD), cadmium (Cd2+), and tetracycline (TC), the removal efficiencies were 98.69 %, 83.95 %, 97.26 %, 98.34 %, and 69.12 % at a hydraulic retention time (HRT) of 4 h, pH of 7.0, and influent salinity, Ca2+, and TC concentrations of 1.0, 180.0, and 3.0 mg/L, respectively. The biofilm reactor packing has a three-dimensional structure to ensure good loading of microorganisms while promoting electron transfer and metabolic activity of microorganisms and increasing the pollutant tolerance and removal efficiency. The reactor provides a practical reference for the sedimentation of reverse osmosis concentrate to remove Cd2+ and TC by microbial induced calcium precipitation (MICP).

Fructose-bisphosphatase1 (FBP1) alleviates experimental osteoarthritis by regulating Protein crumbs homolog 3 (CRB3).

PURPOSE: To identify the role of gluconeogenesis in chondrocytes in osteoarthritis (OA). MATERIALS AND METHODS: Cartilage samples were collected from OA patients and C57 mice and were stained with Safranin O-Fast Green to determine the severity of OA. Periodic acid Schiff staining was used to characterize the contents of polysaccharides and SA-ßGal staining was used to characterize the aging of chondrocytes. Immunohistochemistry and western blotting were used to detect fructose-bisphosphatase1 (FBP1), SOX9, MMP13, P21, and P16 in cartilage or chondrocyte. The mRNA levels of fbp1, mmp13, sox9, colX, and acan were analyzed by qPCR to evaluate the role of FBP1 in chondrocytes. RESULTS: The level of polysaccharides in cartilage was reduced in OA and the expression of FBP1 was also reduced. We treated the chondrocytes with IL-1ß to cause OA in vitro, and then made chondrocytes overexpress FBP1 with plasma. It shows that FBP1 alleviated the degeneration and senescence of chondrocytes in vitro and that it also showed the same effects in vivo experiments. To further understand the mechanism of FBP1, we screened the downstream protein of FBP1 and found that CRB3 was significantly downregulated. And we confirmed that CRB3 suppressed the degeneration and delayed senescence of chondrocytes. CONCLUSIONS: FBP1 promoted the polysaccharide synthesis in cartilage and alleviated the degeneration of cartilage by regulating CRB3, so FBP1 is a potential target in treating OA.

Robust Sparse Bayesian Two-Dimensional Direction-of-Arrival Estimation with Gain-Phase Errors.

To reduce the influence of gain-phase errors and improve the performance of direction-of-arrival (DOA) estimation, a robust sparse Bayesian two-dimensional (2D) DOA estimation method with gain-phase errors is proposed for L-shaped sensor arrays. The proposed method introduces an auxiliary angle to transform the 2D DOA estimation problem into two 1D angle estimation problems. A sparse representation model with gain-phase errors is constructed using the diagonal element vector of the cross-correlation covariance matrix of two submatrices of the L-shaped sensor array. The expectation maximization algorithm derives unknown parameter expression, which is used for iterative operations to obtain off-grid and signal precision. Using these parameters, a new spatial spectral function is constructed to estimate the auxiliary angle. The obtained auxiliary angle is substituted into a sparse representation model with gain and phase errors, and then the sparse Bayesian learning method is used to estimate the elevation angle of the incident signal. Finally, according to the relationship of the three angles, the azimuth angle can be estimated. The simulation results show that the proposed method can effectively realize the automatic matching of the azimuth and elevation angles of the incident signal, and improves the accuracy of DOA estimation and angular resolution.

Effects of Manufactured Sand and Steam-Curing Temperature on the Compressive Strength of Recycled Concrete with Different Water/Binder Ratios.

New building materials (manufactured sand and recycled coarse aggregates) can conserve raw materials and protect the environment. Prefabricated members can shorten the construction time of a structure. To use manufactured sand and recycled coarse aggregate in the preparation of precast member concrete, an economical and practical steam-curing scheme must be developed such that the compressive strength of precast manufactured sand recycled concrete (MRC) meets the requirements for hoisting. The effects of different steam-curing temperatures (standard curing, 40 °C, 50 °C, 60 °C, 70 °C, and 80 °C) on the compressive strength of MRC with three water/binder ratios (W/B) (0.46, 0.42, and 0.38) were studied. In addition, the microstructure of MRC was examined using a scanning electron microscope. The equivalent age-compressive strength model was used to estimate the recycled concrete with manufactured sand. The results showed that the strength of MRC with a water-cement ratio of 0.46, 0.42, and 0.38 reached 33.9, 38.7, and 45.1 MPa, respectively, after 28 days of standard curing. The results also indicated that an increase in the steam-curing temperature had a positive effect on the early compressive strength of MRC and a negative effect on the 28 d compressive strength. This behavior was more obvious for MRC with a low W/B ratio. For MRC with a W/B of 0.46, 0.42, and 0.38, after steam-curing for 6 h, the compressive strength reached 32-65%, 36-70%, and 40-77% of the design strength, respectively. The optimum steam-curing temperatures for MRC with W/B of 0.46, 0.42, and 0.38 were 60 °C, 60 °C, and 50 °C. A decrease in W/B has a negative impact on the accuracy of MRC estimation using the equivalent age-compressive strength model. The maximum deviation of the prediction was within 10%, and the accuracy of the model was acceptable. This study provides a useful reference for the production of prefabricated MRC components in factories and subsequent construction.

Characteristics of the complete chloroplast genome of Swertia divaricata Harry Sm. (Gentianaceae) and its phylogenetic inference.

Swertia divaricata Harry Sm., 1965, (Gentianaceae) is a perennial herb endemic to Northwest Yunnan, China, belonging to the species-rich genus Swertia. It possesses unique morphological features but its systematic position remains uncertain. To determine its phylogenetic placement, the complete plastid genome of S. divaricata was assembled utilizing high-throughput sequencing data. The genome is circular, spanning 152,073 bp, and comprises a large single-copy (LSC) region of 82,470 bp, a small single-copy (SSC) region of 18,153 bp, and two inverted repeats (IR) regions, each 25,725 bp. A total of 130 genes were annotated, including 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. The plastome of S. divaricata exhibits a structure and gene composition highly similar to those of other Swertia plastomes. Phylogenetic analysis indicated that S. divaricata is closely related to S. erythrosticta, sister to a subclade comprising species from sections Swertia and Apterae. The plastome sequence described herein constitutes a valuable contribution to phylogenetic and evolutionary research on Swertia.

Effect of CoSn3 nanocrystals on Sn3Ag plating for electronic packaging.

Plating Sn3Ag on copper substrates represents a crucial electronic packaging technique. In this study, we propose a novel composite plating approach, wherein CoSn3 nanocrystals are deposited within the Sn3Ag coating. The resulting reflowed Sn3Ag joints exhibit a range of distinctive properties. Notably, CoSn3 nanocrystals dissolve in Sn during the reflow process, thereby lowering the supercooling required for Sn nucleation. Consequently, Sn crystals grow in six-fold cyclic twins. Additionally, the dissolution of Co atoms in Sn leads to a reduced solubility of Cu atoms in Sn, consequently lowering the supercooling required for the nucleation of Cu6Sn5. Simultaneously, this phenomenon promotes the nucleation of Cu6Sn5, resulting in a considerable precipitation of Cu6Sn5 nanoparticles within the joints. Therefore, the mechanical properties of the joints are significantly enhanced, leading to a notable 20% increase in shear strength. Furthermore, the presence and distribution of Co elements within Sn induce changes in the growth pattern of interfacial Cu6Sn5. The growth process of Cu6Sn5 is dominated by the interfacial reaction, leading to its growth in a faceted shape. During the aging process, the dissolution of Co elements in Sn impedes the continuous growth of Cu6Sn5 at the interface, causing Cu6Sn5 to be distributed in the form of islands inside the joint. Remarkably, elemental Co acts as an inhibitor for the development of Cu3Sn and reduces the occurrence of Kirkendall voids.

Intra-articular Histone Deacetylase Inhibitor Microcarrier Delivery to Reduce Osteoarthritis.

The histone deacetylase inhibitor (HDACi) was a milestone in the treatment of refractory T-cell lymphoma. However, the beneficial effects of HDACi have not been appreciated in osteoarthritis (OA). Herein, we implemented a microcarrier system because of the outstanding advantages of controlled and sustained release, biodegradability, and biocompatibility. The poly(d,l-lactide-co-glycolide) (PLGA) microcapsules have a regulated and sustained release profile with a reduced initial burst release, which can improve the encapsulation efficiency of the Chidamide. The emulsion solvent evaporation strategy was used to encapsulate Chidamide in PLGA microcapsules. The encapsulation of Chidamide was established by UV-vis spectra and scanning electron microscopy. Additionally, the inhibition of Tnnt3 and immune stimulation by Chidamide helped to inhibit cartilage destruction and prevent articular cartilage degeneration. Based on the results, the Chidamide in PLGA microcapsules provides a transformative therapeutic strategy for the treatment of osteoarthritis patients to relieve symptoms and protect against cartilage degeneration.

Can Platelets/Mean Platelet Volume Accurately Diagnose Periprosthetic Joint Infection? Revealing Their Actual Diagnostic Efficacy.

Background: Currently, there is no single test indicator for diagnosing periprosthetic joint infection (PJI) with an acceptable level of sensitivity. Therefore, ratio indicators have been introduced to improve the accuracy of diagnostic algorithms. Platelet count /mean platelet volume (PMR) is reported to be a potential PJI diagnostic biomarker, but its clinical value for diagnosing PJI is still uncertain. This study aims to provide additional evidence to support the effectiveness of PMR in accurately diagnosing PJI. Methods: This study recruited 116 patients with PJI and 137 patients with aseptic loosening, divided them into PJI group and AL group. Collect subjects' preoperative laboratory indicators such as ESR, CRP, PLT, MPV, etc. The area under the curve (AUC) was calculated by plotting the receiver operating characteristic (ROC) curve to determine the diagnostic efficacy of PMR. Results: ESR, CRP, PLT, and PLT/MPV were significantly increased in the PJI group, while MPV levels were decreased (both P< 0.001). The AUC of the PMR was 0.752, and the optimal cut-off value for diagnosing chronic PJI was determined to be 27.8 based on the Youden index. The sensitivity and specificity for diagnosing PJI were 79.3% and 47.9%, respectively, with a positive predictive value of 68.27%, a negative predictive value of 69.80%, and a diagnostic odds ratio of 4.97. The AUC (0.752) of the ratio biomarker was lower than that of ESR (0.825) and CRP (0.900). After predictive model calculation, the combination of PMR, CRP, and ESR had an AUC value of 0.910, with a sensitivity of 84.5% and a specificity of 84.7%, showing good discriminative ability. Conclusion: Compared with traditional biomarkers ESR and CRP, the value of the PMR for diagnosing PJI is not significant, but it can be used as an auxiliary indicator for PJI diagnosis in combination with other indicators (P<0.001).

Real-Time Evaluation of Helicobacter pylori Infection by Convolution Neural Network During White-Light Endoscopy: A Prospective, Multicenter Study (With Video).

INTRODUCTION: Convolutional neural network during endoscopy may facilitate evaluation of Helicobacter pylori infection without obtaining gastric biopsies. The aim of the study was to evaluate the diagnosis accuracy of a computer-aided decision support system for H. pylori infection (CADSS-HP) based on convolutional neural network under white-light endoscopy. METHODS: Archived video recordings of upper endoscopy with white-light examinations performed at Sir Run Run Shaw Hospital (January 2019-September 2020) were used to develop CADSS-HP. Patients receiving endoscopy were prospectively enrolled (August 2021-August 2022) from 3 centers to calculate the diagnostic property. Accuracy of CADSS-HP for H. pylori infection was also compared with endoscopic impression, urea breath test (URT), and histopathology. H. pylori infection was defined by positive test on histopathology and/or URT. RESULTS: Video recordings of 599 patients who received endoscopy were used to develop CADSS-HP. Subsequently, 456 patients participated in the prospective evaluation including 189 (41.4%) with H. pylori infection. With a threshold of 0.5, CADSS-HP achieved an area under the curve of 0.95 (95% confidence interval [CI], 0.93-0.97) with sensitivity and specificity of 91.5% (95% CI 86.4%-94.9%) and 88.8% (95% CI 84.2%-92.2%), respectively. CADSS-HP demonstrated higher sensitivity (91.5% vs 78.3%; mean difference = 13.2%, 95% CI 5.7%-20.7%) and accuracy (89.9% vs 83.8%, mean difference = 6.1%, 95% CI 1.6%-10.7%) compared with endoscopic diagnosis by endoscopists. Sensitivity of CADSS-HP in diagnosing H. pylori was comparable with URT (91.5% vs 95.2%; mean difference = 3.7%, 95% CI -1.8% to 9.4%), better than histopathology (91.5% vs 82.0%; mean difference = 9.5%, 95% CI 2.3%-16.8%). DISCUSSION: CADSS-HP achieved high sensitivity in the diagnosis of H. pylori infection in the real-time test, outperforming endoscopic diagnosis by endoscopists and comparable with URT. Clinicaltrials.gov ; ChiCTR2000030724.

Design and Control of a Trapezoidal Piezoelectric Bimorph Actuator for Optical Fiber Alignment.

To align a pair of optical fibers, it is required that the micro actuators used be small and have the characteristics of high accuracy and fast response time. A trapezoidal piezoelectric bimorph actuator was proposed for pushing and pulling an optical fiber. Based on a mathematical model and finite element model established in this paper, we analyzed the output displacement and output force of the proposed trapezoidal piezoelectric actuator under the influence of structural parameters. Since the piezoelectric bimorph actuator had a hysteresis effect, we applied particle swarm optimization to establish a Prandtl-Ishlinskii (PI) model for actuator and parameter identification. Then, two control methods, namely feedforward control considering hysteresis effects and fuzzy proportional-integral-derivative (PID) control employing feedback, were proposed. Finally, a composite control model combining the two control methods with fewer tracking errors was designed. The results show that the output displacement of this actuator is larger than that of a rectangular one. Additionally, the fuzzy PID control has a lower response time (15 ms) and an overshoot (5%).

18F-FDG PET/CT in a Patient With Epididymo-Testicular Malacoplakia.

ABSTRACT: A 56-year-old man presented with a 2-month history of a mass in the right epididymo-testicular region, which exhibited heterogeneous high avidity for 18F-FDG on PET/CT. Malignant tumor was highly suspected, leading to subsequent right orchiectomy and epididymectomy. Histopathological examination revealed the presence of characteristic Michaelis-Gutmann bodies within von Hansemann macrophages, confirming the diagnosis of malacoplakia.

A Cas12a-based fluorescent microfluidic system for rapid on-site human papillomavirus diagnostics.

Persistent infection with human papillomavirus (HPV) is the leading cause of cervical cancer, and early diagnosis is crucial for clinical management. However, the easy and rapid on-site diagnostic for HPV genotyping remains challenging. Here, we develop a Cas12a-based fluorescent microfluidic detection system for diagnosing six HPV subtypes (HPV6, HPV11, HPV16, HPV18, HPV31, and HPV33). A panel of crRNAs and recombinase polymerase amplification (RPA) primers targeting the HPV L1 gene was screened for sensitive and specific detection. Furthermore, a one-pot RPA reaction was developed to amplify the six HPV subtypes without cross-reactivity. For on-site detection, we integrated the RPA-Cas12a detection into a microfluidic device, enabling the detection of processed clinical samples within 35 minutes. The assay was validated using 112 clinical swab samples and obtained consistent results with the qPCR assay, with a concordance rate of 99.1%. Overall, our diagnostic method offers a rapid, sensitive, and easy-to-use on-site assay for detecting HPV genotypes and holds promise for improving cervical cancer screening and prevention. KEY POINTS: • The Cas12a-based fluorescent microfluidic detection system for the diagnosis of six HPV subtypes. • A one-pot RPA reaction for amplifying the six HPV subtypes without cross-reactivity. • The RPA-Cas12a-microfluidic system provides results within 35 minutes for on-site detection.

Customized Trajectory Optimization and Compliant Tracking Control for Passive Upper Limb Rehabilitation.

Passive rehabilitation training in the early poststroke period can promote the reshaping of the nervous system. The trajectory should integrate the physicians' experience and the patient's characteristics. And the training should have high accuracy on the premise of safety. Therefore, trajectory customization, optimization, and tracking control algorithms are conducted based on a new upper limb rehabilitation robot. First, joint friction and initial load were identified and compensated. The admittance algorithm was used to realize the trajectory customization. Second, the improved butterfly optimization algorithm (BOA) was used to optimize the nonuniform rational B-spline fitting curve (NURBS). Then, a variable gain control strategy is designed, which enables the robot to track the trajectory well with small human-robot interaction (HRI) forces and to comply with a large HRI force to ensure safety. Regarding the return motion, an error subdivision method is designed to slow the return movement. The results showed that the customization force is less than 6 N. The trajectory tracking error is within 12 mm without a large HRI force. The control gain starts to decrease in 0.5 s periods while there is a large HRI force, thereby improving safety. With the decrease in HRI force, the real position can return to the desired trajectory slowly, which makes the patient feel comfortable.