Differentiating Microplastics from Natural Particles in Aqueous Suspensions Using Flow Cytometry with Machine Learning.

Microplastics (MPs) in natural waters are heterogeneously mixed with other natural particles including algal cells and suspended sediments. An easy-to-use and rapid method for directly measuring and distinguishing MPs from other naturally present colloids in the environment would expedite analytical workflows. Here, we established a database of MP scattering and fluorescence properties, either alone or in mixtures with natural particles, by stain-free flow cytometry. The resulting high-dimensional data were analyzed using machine learning approaches, either unsupervised (e.g., viSNE) or supervised (e.g., random forest algorithms). We assessed our approach in identifying and quantifying model MPs of diverse sizes, morphologies, and polymer compositions in various suspensions including phototrophic microorganisms, suspended biofilms, mineral particles, and sediment. We could precisely quantify MPs in microbial phototrophs and natural sediments with high organic carbon by both machine learning models (identification accuracies over 93%), although it was not possible to distinguish between different MP sizes or polymer compositions. By testing the resulting method in environmental samples through spiking MPs into freshwater samples, we further highlight the applicability of the method to be used as a rapid screening tool for MPs. Collectively, this workflow can be easily applied to a diverse set of samples to assess the presence of MPs in a time-efficient manner.

3D spheroid HepaRG and fluorescent biphasic tracer for CYP3A4-mediated antibiotic interaction monitoring in sepsis.

Cytochrome P450 3A4 (CYP3A4) is a crucial enzyme in the metabolism of xenobiotics, particularly in drug metabolism interactions (DDIs), making it a significant factor in clinical drug use. However, current assay techniques are both laborious and costly, making it difficult to construct a high-throughput monitoring method that can be used in conjunction with the clinic. This poses certain safety hazards for drug combination. Therefore, it is crucial to develop a synchronized monitoring method for the inhibition and induction of CYP3A4. In this study, we utilized 3D culture technology to develop a HepaRG cells spheroid model. The CYP450 and transporter expression, the albumin secretion, and urea synthesis capacity characteristics were analyzed. The NEN probe was utilized as a tracer molecule for CYP3A4. The fluorescence intensity of metabolites was characterized by laser confocal technique to determine the inhibition and expression of CYP3A4 in the HepaRG cell spheroid model by the antibiotics for sepsis. The results indicate that the HepaRG sphere model successfully possessed the physiological phenotype of the liver, which could be used for drug interaction monitoring. Through positive drug testing, NEN probe was able to achieve bidirectional characterization of CYP3A4 induction and inhibition. The monitoring method described in this paper was successfully applied to drug interaction monitoring of commonly used antibiotics in sepsis patients, which is a convenient and rapid monitoring method. The proposed method offers a new strategy for monitoring CYP3A4-mediated drug-drug interactions with a high-throughput assay, which will help to improve the safety of clinical drug combination.

A rapid and multi-endpoint ecotoxicological test using Mychonastes afer for efficient screening of metals and herbicides.

Microalgal growth-based tests are international standards for ecotoxicity assessment; however, their long exposure times, large sample volumes, and reliance on a single growth-endpoint make them inadequate for rapid toxicity screening. Here, we aimed to develop a rapid and simple ecotoxicological test using the fast-growing green alga Mychonastes afer, with multiple endpoints-growth, lipid content, and photosynthesis. We exposed M. afer to two metals-silver and copper-and two herbicides-atrazine and diuron-for 24 h and identified the most sensitive and reliable endpoints for each toxicant: the maximum electron transport rate (ETRmax) for Ag, Cu and atrazine, and the lipid content for diuron. Lipid content was found to be both a sensitive and reliable biomarker, meeting the effluent limit guidelines in both the Republic of Korea and the USA. The sensitivity of M. afer to Ag and atrazine also closely matched the HC5 values derived from the species sensitivity distribution approach, confirming its reliability for setting regulatory concentrations of these contaminants. Our calculated predicted no-effect concentration (PNEC) values were similar to established European Union PNECs for Ag, Cu, atrazine, and diuron, underlining the utility of these biological endpoints for ecological risk assessment and regulatory decision making. This method required lower sample volume (2 mL vs 100 mL) and exposure time (24 h vs 72-120 h) than conventional green algal tests, and eliminated the need for labour-intensive cell counting, expensive equipment, and chlorophyll fluorescence measurement expertise. Overall, this M. afer test can be a valuable tool for the rapid screening of wastewater for metals and herbicides, contributing to environmental protection and management practices.

Discriminating extra virgin olive oils from common edible oils: Comparable performance of PLS-DA models trained on low-field and high-field 1H NMR data.

INTRODUCTION: Olive oil, derived from the olive tree (Olea europaea L.), is used in cooking, cosmetics, and soap production. Due to its high value, some producers adulterate olive oil with cheaper edible oils or fraudulently mislabel oils as olive to increase profitability. Adulterated products can cause allergic reactions in sensitive individuals and can lack compounds which contribute to the perceived health benefits of olive oil, and its corresponding premium price. OBJECTIVE: There is a need for robust methods to rapidly authenticate olive oils. By utilising machine learning models trained on the nuclear magnetic resonance (NMR) spectra of known olive oil and edible oils, samples can be classified as olive and authenticated. While high-field NMRs are commonly used for their superior resolution and sensitivity, they are generally prohibitively expensive to purchase and operate for routine screening purposes. Low-field benchtop NMR presents an affordable alternative. METHODS: We compared the predictive performance of partial least squares discrimination analysis (PLS-DA) models trained on low-field 60 MHz benchtop proton (1H) NMR and high-field 400 MHz 1H NMR spectra. The data were acquired from a sample set consisting of 49 extra virgin olive oils (EVOOs) and 45 other edible oils. RESULTS: We demonstrate that PLS-DA models trained on low-field NMR spectra are highly predictive when classifying EVOOs from other oils and perform comparably to those trained on high-field spectra. We demonstrated that variance was primarily driven by regions of the spectra arising from olefinic protons and ester protons from unsaturated fatty acids in models derived from data at both field strengths.

A RaPID Macrocyclic Peptide That Inhibits the Formation of α-Synuclein Amyloid Fibrils.

There is considerable interest in drug discovery targeting the aggregation of α-synuclein (αSyn) since this molecular process is closely associated with Parkinson's disease. However, inhibiting αSyn aggregation remains a major challenge because of its highly dynamic nature which makes it difficult to form a stable binding complex with a drug molecule. Here, by exploiting Random non-standard Peptides Integrated Discovery (RaPID) system, we identified a macrocyclic peptide, BD1, that could interact with immobilized αSyn and inhibit the formation of fibrils. Furthermore, improving the solubility of BD1 suppresses the co-aggregation with αSyn fibrils while it kinetically inhibits more effectively without change in their morphology. We also revealed the molecular mechanism of kinetic inhibition, where peptides bind to fibril ends of αSyn, thereby preventing further growth of fibrils. These results suggest that our approach for generating non-standard macrocyclic peptides is a promising approach for developing potential therapeutics against neurodegeneration.

Plastic probe electrospray ionization mass spectrometry developed for rapid fingerprint profile of biological samples without pretreatment.

A triangular-shaped flat plastic substrate probe was prepared for direct electrospray ionization mass spectrometry (ESI-MS) for analysis of untreated chemical and biological samples including liquids (Met-Arg-Phe-Ala peptide, reserpine, and dodecyl aldehyde), solids (biological samples, traditional Chinese medicine), and powders (roasted coffee, rhizoma coptidis, lotus plumule, and Schisandra sphenanthera). Quantitative analysis of reserpine in water yielded a detection limit of 1 ng mL-1, dynamic response range within 1-500 ng mL-1, and linearity of signal response ˃0.9925. Compared to the conventional capillary ESI, this plastic probe ESI offers lower cost of analysis (US $0.0056 per probe), higher sensitivity, lower sample consumption, longer signal duration (>6 min), better reproducibility, signal stability, and higher speed of analysis (<10 s per sample, including sample loading). Overall, the results indicate the potential of ESI-MS based on flat plastic probes as a versatile method for simple, sensitive, and stable analysis of untreated biological sample analysis.

Metrological traceability in process analytical technologies and point-of-need technologies for food safety and quality control: not a straightforward issue.

Traditional techniques for food analysis are based on off-line laboratory methods that are expensive and time-consuming and often require qualified personnel. Despite the high standards of accuracy and metrological traceability, these well-established methods do not facilitate real-time process monitoring and timely on-site decision-making as required for food safety and quality control. The future of food testing includes rapid, cost-effective, portable, and simple methods for both qualitative screening and quantification of food contaminants, as well as continuous, real-time measurement in production lines. Process automatization through process analytical technologies (PAT) is an increasing trend in the food industry as a way to achieve improved product quality, safety, and consistency, reduced production cycle times, minimal product waste or reworks, and the possibility for real-time product release. Novel methods of analysis for point-of-need (PON) screening could greatly improve food testing by allowing non-experts, such as consumers, to test in situ food products using portable instruments, smartphones, or even visual naked-eye inspections, or farmers and small producers to monitor products in the field. This requires the attention of the research community and devices manufacturers to ensure reliability of measurement results from PAT strategy and PON tests through the demonstration and critical evaluation of performance characteristics. The fitness for purpose of methods in real-life conditions is a priority that should not be overlooked in order to maintain an effective and harmonized food safety policy.

Rapid screening of polybrominated diphenyl ethers in water by solid-phase microextraction coupled with ultrahigh-resolution mass spectrometry.

Polybrominated diphenyl ethers (PBDEs) are considered emerging organic contaminants that attract more attention in the environment. Herein, online coupling of solid-phase microextraction and ultrahigh-resolution mass spectrometry was developed for rapid screening of eight PBDEs in water samples. This procedure was completed in 22 min, about 6 times faster than the routine workflow such as solid-phase extraction coupled with gas chromatography-mass spectrometry. Thermal desorption and solvent-assisted atmospheric pressure chemical ionization were developed for the effective coupling of solid-phase microextraction (SPME) with ultrahigh-resolution mass spectrometry (UHRMS), which contributed to the signal enhancement and made the methodology feasible for environmental screening. The limits of detection and quantification were 0.01-0.50 ng/mL and 0.05-4.00 ng/mL, respectively. The recoveries were 57.2-75.2% for quality control samples at spiking levels of 0.8-10 ng/mL (4-50 ng/mL for BDE209), with relative standard deviation less than 19.0%. Twelve water samples from different river sites near industrial areas were screened using the developed method. The results showed that BDE-209 was the dominant PBDE (1.02-1.28 ng/mL in positive samples), but its amount was lower than the human health ambient water quality criteria. Consequently, the developed method provides a rapid and reliable way of evaluating contamination status and risks of PBDEs in aqueous environment.

Screening Method and Antibacterial Activity of 1,3,4-Oxadiazole Sulfone Compounds against Citrus Huanglongbing.

Citrus Huanglongbing (HLB) is one of the most destructive diseases in the citrus industry. At present, Candidatus Liberibacter asiaticus (CLas) cannot be cultured in vitro, and there is a lack of rapid methods to test antibacterial activity, which greatly hinders the discovery of new antibacterial agents against HLB. To establish a rapid screening method for antibacterial agents against HLB with simple operation, a short cycle, and a large number of tests, the CLas contents in leaves from different citrus branches, different leaves from the same citrus branch, and two halves of the same citrus leaf were detected. Compared with the leaves on different branches and different leaves on the same branch, the difference in CLas content of the left and right halves of the same leaf was small; the difference was basically between 0.7 and 1.3. A rapid and efficient method for primary screening agents against HLB termed the "half-leaf method" was established through our long-term optimization and improvement. To verify the stability and reliability of the activity data measured using this method, 6-chloropurine riboside, which is highly soluble in water, was used as the test agent, and its antibacterial activity against HLB was tested 45 times. The results of the antibacterial activity test showed little difference in the mean values of each data group, indicating that this method could be used as a rapid method for screening agents against HLB. We used this method to test the antibacterial activity of compounds synthesized by our research group against HLB and found that some of the compounds showed good activity.

Investigation of Chemical Constituents and Antioxidant Activity of Biologically Active Plant-Derived Natural Products.

The aim of this publication is to present rapid screening methods (visual/colorimetric) that will enable quick identification of the presence of biologically active compounds in aqueous solutions. For this reason, 26 plant extracts obtained by ultrasound-assisted extraction were analysed for the content of these compounds. Higher plants, used as a raw material for extraction, are common in Europe and are easily available. The article proposes a comparison of various protocols for the identification of various compounds, e.g., phenolic compounds (phenols, tannins, anthocyanins, coumarins, flavones, flavonoids), vitamin C, quinones, quinines, resins, glycosides, sugars. Initial characterisation of the composition of plant extracts using fast and inexpensive methods allows you to avoid the use of time-consuming analyses with the use of advanced research equipment. In addition, the antioxidant activity of plant extracts using spectrophotometric methods (DPPH, ABTS, FRAP assay) and quantitative analysis of plant hormones such as abscisic acid, benzoic acid, gibberellic acid, indole acetic acid, jasmonic acid, salicylic acid, zeatin, zeatin riboside, and isipentenyl adenine was performed. The obtained results prove that the applied visual methods show different sensitivity in detecting the sought chemical compounds. Therefore, it is necessary to confirm the presence or absence of bioactive substances and their concentration using modern analytical methods.

Rapid screening based on machine learning and molecular docking of umami peptides from porcine bone.

BACKGROUND: The traditional screening method for umami peptide, extracted from porcine bone, was labor-intensive and time-consuming. In this study, the rapid screening method and molecular mechanism of umami peptide was investigated. RESULTS: This article showed that a more precisely rapid screening method with composite machine learning and molecular docking was used to screen the potential umami peptide from porcine bone. As reference, 24 reported umami peptides were predicated by composite machine learning, with the accuracy of 86.7%. In this study, potential umami peptide sequences from porcine bone were screened by UMPred-FRL, Umami-MRNN Demo, and molecular docking was used to provide further screening. Finally, nine peptides were screened and verified as umami peptides by this method: LREY, HEAL, LAKVH, FQKVVA, HVKELE, AEVKKAP, EAVEKPQS, KALSEEL and KKMFETES. The hydrogen bonding was deemed to be the main interaction force with receptor T1R3, and domain binding sites were Ser146, His121 and Glu277. The result demonstrated the feasibility of machine learning assisted T1R1/T1R3 receptor for rapid screening umami peptides. The screening method would not only adapt to screen umami peptides from porcine bone but possibly applied for other sources. It also provided a reference for rapid screening of umami peptides. CONCLUSION: The manuscript lays a rapid screening method in screening umami peptide, and nine umami peptides from porcine bone were screened and identified. © 2022 Society of Chemical Industry.

Single-Tube Screen for Rapid Detection of Repeat Expansions in Seven Common Spinocerebellar Ataxias.

BACKGROUND: The autosomal dominantly inherited and genetically heterogeneous spinocerebellar ataxias (SCAs) exhibit highly similar clinical presentations. Many are caused by repeat expansions, of which at least 8 involve CAG repeats. Repeat expansion detection is the only method to confirm disease status in symptomatic individuals. We present a novel strategy to simultaneously screen for the presence of CAG repeat expansion in the genes responsible for SCAs 1, 2, 3, 6, 7, 12, and dentatorubral-pallidoluysian atrophy using a simplified single-tube assay. METHODS: The method employs differentially labeled locus-specific primers and a common triplet-primed primer. Amplified products from each locus are distinguished by a combination of the product size and the fluorophore tag. The upper size limit of the normal allele range was used as the cutoff for distinguishing normal from potentially affected samples, with repeat expansion detected by presence of electrophoretic peaks extending beyond the cutoff. RESULTS: Blinded evaluation of the assay on 60 genotype-known DNA samples correctly detected repeat expansion in the expected SCA repeat locus for all 31 DNA samples. CONCLUSIONS: In principle, this strategy can be applied to the simultaneous screening of any group of disease genes sharing the same repetitive units and/or their reverse complement.

Investigating public awareness, prevailing attitudes and perceptions towards domestic violence and abuse in the United Kingdom: a qualitative study.

BACKGROUND: Reported cases of Domestic Violence and Abuse (DVA) have increased since the advent of the COVID-19 pandemic and ensuing lockdowns. Understanding the general public's view about DVA is vital, as it would help develop targeted interventions and effective public policies to tackle this rising problem in society. Our qualitative study investigated the public awareness, attitudes and perceptions towards DVA, and explored mechanisms to tackle DVA in the community setting in the UK. METHODS: The research team conducted personal interviews with 29 community dwelling adults who responded to study invitations and adverts on social media. We used a topic guide to ensure consistency across the interviews, which were audio-recorded, transcribed and analysed thematically to detect emergent themes concerning DVA. RESULTS: All respondents were aware of the concept of abuse. Thirty-eight percent declared either having experienced DVA directly or that they knew someone close to being abused. More than half of the respondents were not aware of existing DVA supportive services in the UK. Overarching themes generated from the contextual analysis included contributing factors for DVA, challenges and barriers facing victims and proposals for future interventions. CONCLUSIONS: Community dwelling adults have a good understanding of the impacts of DVA, but many fail to recognise specific instances or events in their daily lives contributing to DVA. Raising public awareness, particularly in children through the school curriculum, highlighting existing support services and introducing the routine use of short screening tools for DVA in health and social care settings can increase awareness, early identification and signposting to effective interventions. Sustained, multi-level community facing interventions are recommended to reduce stigma and fear associated with DVA.

Rapid and simultaneous visual typing of high-risk HPV-16/18 with use of integrated lateral flow strip platform.

A biosensor for rapid and simultaneous visual identification of high-risk human papillomavirus (HPV) genotypes 16 and 18 in clinical samples based on polymerase chain reaction (PCR) integrated lateral flow strip platform was developed. Using an one-step protocol to extract nucleic acid rapidly and the functionalized primer sets specific to HPV-16 and 18 were designed for the simultaneous amplification. In the presence of target HPV genotypes, the corresponding functionalized primer sets will participate in the PCR process and produce numerous duplex functionalized dsDNA amplicons. With the bridge effect of duplex functionalized dsDNA amplicons between gold nanoparticles-fluorescein isothiocyanate antibody conjugates (AuNP-FITC antibody conjugates) and other two antibodies on corresponding test line (T1 or T2), visualized color signals on test lines could be obtained directly visible with a naked eye. Combining the high amplification efficiency of PCR and the visualized sensing of LFS, as low as 700 copies of HPV-16 and 18 DNA were detected simultaneously within 75 min, which can promote application in the resource limited settings. High-risk genotypes of HPV-16 and HPV-18 were easily and simultaneously screened with the amplification-assisted molecular lateral flow strip by on-site observation in the resource-limited settings.

Prompt successful response to a COVID-19 outbreak: Performance of community-based rapid screening station.

An outbreak occurred in Wanhua District of Taipei City. It was traced to a cluster infection originating from a teahouse. To prevent further large-scaled community spread, the Taipei City Government established the first community rapid test screening station. This report describes the station's strategy and performance and key factors that contributed to its operation. The project involves collaboration among various departments of Taipei City Government, including the health, environmental, police, transportation, and fire departments. The station provides rapid screening, polymerase chain reaction (PCR) testing, and immediate isolation and follow-up medical services upon the detection of a positive case. These services are accessible to local residents and are intended to ease hospitals' burdens. In 36 days, a total of 8532 people were tested, and 419 confirmed cases were identified. Over the same period, the weekly number of positive cases in Wanhua District decreased from 356 to 40, and the PCR positive rate decreased from 21.7% to 1.2%. The policy of establishing rapid screening station, contact tracing and mask wearing policy are key strategies for interrupting chains of transmission of COVID-19. This intervention has become a model for preventing the spread of the epidemic and establishing community rapid screening stations in Taiwan.

A prospective study of the decision tree prediction model for attention deficit hyperactivity disorder in preschool children. / å­¦é¾„å‰å„¿ç«¥æ³¨æ„ç¼ºé™·å¤šåŠ¨éšœç¢å†³ç­–æ ‘é¢„æµ‹æ¨¡åž‹çš„å‰çž»æ€§ç ”ç©¶.

OBJECTIVES: To study the clinical value of attention time combined with behavior scale in the screening of attention deficit hyperactivity disorder (ADHD) in preschool children. METHODS: A total of 200 preschool children with ADHD diagnosed in Fujian Maternal and Child Health Hospital from February 2019 to March 2020 were enrolled as the ADHD group. A total of 200 children who underwent physical examination in the hospital or kindergartens during the same period were enrolled as the control group. Attention time was recorded. Chinese Version of Swanson Nolan and Pelham, Version IV Scale-Parent Form (SNAP-IV) scale was used to evaluate symptoms. With clinical diagnosis as the gold standard, the decision tree analysis was used to evaluate the clinical value of attention time combined with behavior scale in the screening of ADHD. RESULTS: Compared with the control group, the ADHD group had significantly higher scores of SNAP-IV items 1, 4, 7, 8, 10, 11, 14, 15, 16, 18, 20, 21, and 22 (P<0.05) and a significantly shorter attention time (P<0.05). The variables with statistically significant differences between the two groups in univariate analysis were used as independent variables to establish a decision tree model. The accuracy of the model in predicting ADHD was 81%, that in predicting non-ADHD was 69%, and the overall accuracy was 75%, with an area under the ROC curve of 0.816 (95% CI: 0.774-0.857, P<0.001). CONCLUSIONS: The decision tree model for screening ADHD in preschool children based on attention time and assessment results of behavior scale has a high accuracy and can be used for rapid screening of ADHD among children in clinical practice.

Rapid screening of high expressing Escherichia coli colonies using a novel dicistronic-autoinducible system.

BACKGROUND: Identification of high-expressing colonies is one of the main concerns in the upstream process of recombinant protein development. The common method to screen high-producing colonies is SDS-PAGE, a laborious and time-consuming process, which is based on a random and qualitative way. The current study describes the design and development of a rapid screening system composed of a dicistronic expression system containing a reporter (enhanced green fluorescent protein, eGFP), protein model (staphylokinase, SAK), and a self-inducible system containing heat shock protein 27 (Hsp27). RESULTS: Dicistronic-autoinducible system expressed eGFP and SAK successfully in 5-ml and 1-L culture volumes. High expressing colonies were identified during 6 h via fluorescent signals. In addition, the biological activity of the protein model was confirmed semi-quantitatively and quantitatively through radial caseinolytic and chromogenic methods, respectively. There was a direct correlation between eGFP fluorescent intensity and SAK activity. The correlation and linearity of expression between the two genes were respectively confirmed with Pearson correlation and linear regression. Additionally, the precision, limit of detection (LOD), and limit of quantification (LOQ) were determined. The expression of eGFP and SAK was stable during four freeze-thaw cycles. In addition, the developed protocol showed that the transformants can be inoculated directly to the culture, saving time and reducing the error-prone step of colony picking. CONCLUSION: The developed system is applicable for rapid screening of high-expressing colonies in most research laboratories. This system can be investigated for other recombinant proteins expressed in E. coli with a potential capability for automation and use at larger scales.

Colloidal gold-based lateral flow immunoassay with inline cleanup for rapid on-site screening of carbendazim in functional foods.

In this study, for the first time, we propose a sensitive colloidal gold-based lateral flow immunoassay (LFIA) that can be used to detect carbendazim residues in functional foods. The adoption of inline cleanup LFIA strips effectively improved background interference to reduce misjudgment of results. First, the hapten 2-(methylamino)-1H-benzo[d]imidazole-5-carboxylic acid was used to establish the carbendazim immunoassay method. Subsequently, colloidal gold-mAb preparation and LFIA detection conditions were systematically optimized. For root and fruit samples (ginseng, ginger, jujube, and Chinese wolfberry), the designed strips had a cutoff value of 8 ng/mL. For flower and seed samples (chrysanthemum, coix seed, and malt), the cutoff value was 12 ng/mL. Even in a complex matrix, the established LFIA method demonstrates satisfactory sensitivity and anti-interference ability. This method was successfully applied in detection of carbendazim residues in complex functional foods, and the assay results are consistent with those obtained via liquid chromatography-tandem mass spectrometry. In short, the proposed method is fast and sensitive and has strong anti-interference ability. Furthermore, it provides a new technical method highly relevant to the on-site rapid detection of carbendazim residues in complex sample matrix.

Recent advance in the discovery of tyrosinase inhibitors from natural sources via separation methods.

Tyrosinase (TYR) inhibitors are in great demand in the food, cosmetic and medical industrials due to their important roles. Therefore, the discovery of high-quality TYR inhibitors is always pursued. Natural products as one of the most important sources of bioactive compounds discovery have been increasingly used for TYR inhibitors screening. However, due to their complex compositions, it is still a great challenge to rapid screening and identification of biologically active components from them. In recent years, with the help of separation technologies and the affinity and intrinsic activity of target enzymes, two advanced approaches including affinity screening and inhibition profiling showed great promises for a successful screening of bioactive compounds from natural sources. This review summarises the recent progress of separation-based methods for TYR inhibitors screening, with an emphasis on the principle, application, advantage, and drawback of each method along with perspectives in the future development of these screening techniques and screened hit compounds.

A clinical study on bone defect reconstruction and functional recovery in benign bone tumors of the lower extremity, treated by bone marrow mesenchymal stem cell rapid screening-enrichment-composite system.

BACKGROUND: With the development of medical technology, credible options for defect reconstructions after the resection of benign bone tumors of the lower extremities have become a high priority. As the current reconstructive methods commonly used in clinical practice have some flaws, new methods of reconstruction need to be explored. We aimed to prepare a new kind of bioactive scaffold for the repair of bone defects through a stem cell rapid screening-enrichment-composite technology system developed by our team. Furthermore, we aimed to investigate the curative effects of these bioactive scaffolds. METHODS: Firstly, cell count, trypan blue exclusion rate, and ALP staining were used to evaluate changes in enrichment efficiency, cell activity, and osteogenic ability before and after enrichment. Then, the scaffolds were placed under the skin of nude mice to verify their osteogenic effects in vivo. Finally, the scaffolds were used for the reconstruction of bone defects after operations for benign bone tumors in a patient's lower limb. The healing status of the defect site at 1 and 3 months was assessed by X-ray, and the Musculoskeletal Tumor Society (MSTS) score was applied to reflect the recovery of patient limb function. RESULTS: The system effectively enriched stem cells without affecting the activity and osteogenic abilities of the bone marrow mesenchymal stem cells (BMSCs). Meanwhile, the bioactive scaffolds obtained better osteogenic effects. In patients, the active scaffolds showed better bone integration and healing status, and the patients also obtained higher MSTS scores at 1 and 3 months after surgery. CONCLUSION: As a new technique, the rapid screening-enrichment-composite technology of stem cells demonstrates a better therapeutic effect in the reconstruction of bone defects after surgery for benign bone tumors of the lower extremities, which will further improve patient prognosis.