Screening membraneless organelle participants with machine-learning models that integrate multimodal features.

Protein self-assembly is one of the formation mechanisms of biomolecular condensates. However, most phase-separating systems (PS) demand multiple partners in biological conditions. In this study, we divided PS proteins into two groups according to the mechanism by which they undergo PS: PS-Self proteins can self-assemble spontaneously to form droplets, while PS-Part proteins interact with partners to undergo PS. Analysis of the amino acid composition revealed differences in the sequence pattern between the two protein groups. Existing PS predictors, when evaluated on two test protein sets, preferentially predicted self-assembling proteins. Thus, a comprehensive predictor is required. Herein, we propose that properties other than sequence composition can provide crucial information in screening PS proteins. By incorporating phosphorylation frequencies and immunofluorescence image-based droplet-forming propensity with other PS-related features, we built two independent machine-learning models to separately predict the two protein categories. Results of independent testing suggested the superiority of integrating multimodal features. We performed experimental verification on the top-scored proteins DHX9, Ki-67, and NIFK. Their PS behavior in vitro revealed the effectiveness of our models in PS prediction. Further validation on the proteome of membraneless organelles confirmed the ability of our models to identify PS-Part proteins. We implemented a web server named PhaSePred (http://predict.phasep.pro/) that incorporates our two models together with representative PS predictors. PhaSePred displays proteome-level quantiles of different features, thus profiling PS propensity and providing crucial information for identification of candidate proteins.

Diagnosis of Nonalcoholic Fatty Liver Disease via a H2S-Responsive Bioluminescent Probe Combined with Firefly Luciferase mRNA Delivery.

The early detection of nonalcoholic fatty liver disease (NAFLD) through bioluminescent probes is of great significance. However, there remains a challenge to apply them in nontransgenic natural animals due to the lack of exogenous luciferase. To address this issue, we herein report a new strategy for in situ monitoring of endogenous hydrogen sulfide (H2S) in the liver of NAFLD mice by leveraging a H2S-responsive bioluminescent probe (H-Luc) combined with firefly luciferase (fLuc) mRNA delivery. The probe H-Luc was created by installing a H2S recognition moiety, 2,4-dinitrophenol, onto the luciferase substrate (d-luciferin), which is allowed to release cage-free d-luciferin in the presence of H2S via a nucleophilic aromatic substitution reaction. In the meantime, the intracellular luciferase was introduced by lipid nanoparticle (LNP)-mediated fLuc mRNA delivery, rendering it suitable for bioluminescence (BL) imaging in vitro and in vivo. Based on this luciferase-luciferin system, the endogenous H2S could be sensitively and selectively detected in living cells, showing a low limit of detection (LOD) value of 0.72 µM. More importantly, after systematic administration of fLuc mRNA-loaded LNPs in vivo, H-Luc was able to successfully monitor the endogenous H2S levels in the NAFLD mouse model for the first time, displaying a 28-fold higher bioluminescence intensity than that in the liver of normal mice. We believe that this strategy may shed new light on the diagnosis of inflammatory liver disease, further elucidating the roles of H2S.

Visualization of Endogenous Hypochlorite in Drug-Induced Liver Injury Mice via a Bioluminescent Probe Combined with Firefly Luciferase mRNA-Loaded Lipid Nanoparticles.

Drug-induced liver injury (DILI) is a common liver disease with a high rate of morbidity, and its pathogenesis is closely associated with the overproduction of highly reactive hypochlorite (ClO-) in the liver. However, bioluminescence imaging of endogenous hypochlorite in nontransgenic natural mice remains challenging. Herein, to address this issue, we report a strategy for imaging ClO- in living cells and DILI mice by harnessing a bioluminescent probe formylhydrazine luciferin (ClO-Luc) combined with firefly luciferase (fLuc) mRNA-loaded lipid nanoparticles (LNPs). LNPs could efficiently deliver fLuc mRNA into living cells and in vivo, expressing abundant luciferase in the cytoplasm in situ. In the presence of ClO-, probe ClO-Luc locked by formylhydrazine could release cage-free d-luciferin through oxidation and follow-up hydrolysis reactions, further allowing for bioluminescence imaging. Moreover, based on the luciferase-luciferin system, it was able to sensitively and selectively detect ClO- in vitro with a limit of detection of 0.59 µM and successfully monitor the endogenous hypochlorite generation in the DILI mouse model for the first time. We postulate that this work provides a new method to elucidate the roles of ClO- in related diseases via bioluminescence imaging.

Assessment and machine learning prediction of heavy metals fate in mining farmland assisted by Positive Matrix Factorization.

The accurate pollutant prediction by Machine Learning (ML) is significant to efficient environmental monitoring and risk assessment. However, application of ML in soil is under studied. In this study, a Positive Matrix Factorization (PMF) assisted prediction method was developed with Support Vector Machine (SVM) and Random Forest (RF) for heavy metals (HMs) prediction in mining farmland. Principal Component Analysis (PCA) and Redundancy Analysis (RDA) were selected to pretreat data. Experiment results illustrated Cd was the main pollutant with heavy risks in the study area and Pb was easy to migrate. The method effects of HMs total concentration predicting were PMF > Simple > PCA > PCA - PMF, and RF predicted better than SVM. Data pretreatment by RDA prior inspection improved the model results. Characteristic HMs Tessier fractions prediction received good effects with average R value as 0.86. Risk classification prediction performed good in Cd, Cu, Ni and Zn, however, Pb showed weak effect by simple model. The best classifier method for Pb was PMF - RF method with relatively good effect (Area under ROC Curve = 0.896). Overall, our study suggested the combination between PMF and ML can assist the prediction of HMs in soil. Spatial weighted attribute of HMs can be provided by PMF.

Endothelial miR-196b-5p regulates angiogenesis via the hypoxia/miR-196b-5p/HMGA2/HIF1α loop.

Angiogenesis is involved in development, reproduction, wound healing, homeostasis, and other pathophysiological events. Imbalanced angiogenesis predisposes patients to various pathological processes, such as angiocardiopathy, inflammation, and tumorigenesis. MicroRNAs (miRNAs) have been found to be important in regulating cellular processing and physiological events including angiogenesis. However, the role of miRNAs that regulate angiogenesis (angiomiRs) is not fully understood. Here, we observed a downregulation of the miR-196 family in endothelial cells upon hypoxia. Functionally, miR-196b-5p inhibited the angiogenic functions of endothelial cells in vitro and suppressed angiogenesis in Matrigel plugs and skin wound healing in vivo. Mechanistically, miR-196b-5p bound onto the 3' untranslated region (UTR) of high-mobility group AT-hook 2 (HMGA2) mRNA and repressed the translation of HMGA2, which in turn represses HIF1α accumulation in endothelial cells upon hypoxia. Together, our results establish the role of endothelial miR-196b-5p as an angiomiR that negatively regulates endothelial growth in angiogenesis via the hypoxia/miR-196b-5p/HMGA2/HIF1α loop. miR-196b-5p and its regulatory loop could be an important addition to the molecular mechanisms underlying angiogenesis and may serve as potential targets for antiangiogenic therapy.

Modular Design of Biodegradable Ionizable Lipids for Improved mRNA Delivery and Precise Cancer Metastasis Delineation In Vivo.

Lipid nanoparticles (LNPs) represent the most clinically advanced nonviral mRNA delivery vehicles; however, the full potential of the LNP platform is greatly hampered by inadequate endosomal escape capability. Herein, we rationally introduce a disulfide bond-bridged ester linker to modularly synthesize a library of 96 linker-degradable ionizable lipids (LDILs) for improved mRNA delivery in vivo. The top-performing LDILs are composed of one 4A3 amino headgroup, four disulfide bond-bridged linkers, and four 10-carbon tail chains, whose unique GSH-responsive cone-shaped architectures endow optimized 4A3-SCC-10 and 4A3-SCC-PH lipids with superior endosomal escape and rapid mRNA release abilities, outperforming their parent lipids 4A3-SC-10/PH without a disulfide bond and control lipids 4A3-SSC-10/PH with a disulfide bond in the tail. Notably, compared to DLin-MC3-DMA via systematic administration, 4A3-SCC-10- and 4A3-SCC-PH-formulated LNPs significantly improved mRNA delivery in livers by 87-fold and 176-fold, respectively. Moreover, 4A3-SCC-PH LNPs enabled the highly efficient gene editing of 99% hepatocytes at a low Cre mRNA dose in tdTomato mice following intravenous administration. Meanwhile, 4A3-SCC-PH LNPs were able to selectively deliver firefly luciferase mRNA and facilitate luciferase expression in tumor cells after intraperitoneal injection, further improving cancer metastasis delineation and surgery via bioluminescence imaging. We envision that the chemistry adopted here can be further extended to develop new biodegradable ionizable lipids for broad applications such as gene editing and cancer immunotherapy.

Analysis of B Cell Receptor Repertoires Reveals Key Signatures of the Systemic B Cell Response after SARS-CoV-2 Infection.

A comprehensive study of the B cell response against SARS-CoV-2 could be significant for understanding the immune response and developing therapeutical antibodies and vaccines. To define the dynamics and characteristics of the antibody repertoire following SARS-CoV-2 infection, we analyzed the mRNA transcripts of immunoglobulin heavy chain (IgH) repertoires of 24 peripheral blood samples collected between 3 and 111 days after symptom onset from 10 COVID-19 patients. Massive clonal expansion of naive B cells with limited somatic hypermutation (SHM) was observed in the second week after symptom onset. The proportion of low-SHM IgG clones strongly correlated with spike-specific IgG antibody titers, highlighting the significant activation of naive B cells in response to a novel virus infection. The antibody isotype switching landscape showed a transient IgA surge in the first week after symptom onset, followed by a sustained IgG elevation that lasted for at least 3 months. SARS-CoV-2 infection elicited poly-germ line reactive antibody responses. Interestingly, 17 different IGHV germ line genes recombined with IGHJ6 showed significant clonal expansion. By comparing the IgH repertoires that we sequenced with the 774 reported SARS-CoV-2-reactive monoclonal antibodies (MAbs), 13 shared spike-specific IgH clusters were found. These shared spike-specific IgH clusters are derived from the same lineage of several recently published neutralizing MAbs, including CC12.1, CC12.3, C102, REGN10977, and 4A8. Furthermore, identical spike-specific IgH sequences were found in different COVID-19 patients, suggesting a highly convergent antibody response to SARS-CoV-2. Our analysis based on sequencing antibody repertoires from different individuals revealed key signatures of the systemic B cell response induced by SARS-CoV-2 infection. IMPORTANCE Although the canonical delineation of serum antibody responses following SARS-CoV-2 infection has been well established, the dynamics of antibody repertoire at the mRNA transcriptional level has not been well understood, especially the correlation between serum antibody titers and the antibody mRNA transcripts. In this study, we analyzed the IgH transcripts and characterized the B cell clonal expansion and differentiation, isotype switching, and somatic hypermutation in COVID-19 patients. This study provided insights at the repertoire level for the B cell response after SARS-CoV-2 infection.

Source identification and health risk assessment of heavy metals with mineralogy: the case of soils from a Chinese industrial and mining city.

Understanding the precise sources of heavy metals (HMs) in soil and the contribution of these sources to health risks has positive effects in terms of risk management. This study focused on the HMs in the soil of five land uses in an industrial and mining city. The sources of HMs in soils were identified, and the soil mineralogical characteristics and health risks of HMs were discussed. The results showed that the HMs (Cu, Zn, Ni, Cd, Pb) found in the soil of the five land uses were affected by human activities. For example, the Cu in grassland, gobi beach, woodland, green belt, and farmland is 22.3, 3.5, 22.5, 16.7, and 21.3 times higher than the soil background values in Gansu Province, respectively. The Positive Matrix Factorization model (PMF) results revealed that traffic emissions and industrial and agricultural activities were the primary sources of HMs in the soil, with industrial sources accounting for the largest share at 55.79%. Furthermore, various characteristics proved that the studied HMs were closely related to smelting products. Concentration-oriented health risk assessments showed that HMs in the different soil types held non-carcinogenic and carcinogenic risks for children and adults. Contamination source-oriented health risk assessments of children and adults found that industrial activities controlled non-carcinogenic and carcinogenic risks. This study highlighted the critical effects of smelting on urban soil and the contribution of pollution sources to health risks. Furthermore, this work is significant in respect of the risk control of HMs in urban soils.

"Dual-Key-and-Lock" NIR-II NSCyanines Enable High-Contrast Activatable Phototheranostics in Extrahepatic Diseases.

Conventional cyanine dyes with a symmetric structure are "always-on", which can easily accumulate in the liver and display high liver background fluorescence, inevitably interfering the accurate diagnosis and therapy in extrahepatic diseases. We herein report a platform of NIR-II non-symmetric cyanine (NSCyanine) dyes by harnessing a non-symmetric strategy, which are extremely sensitive to pH/viscosity and can be activated via a "dual-key-and-lock" strategy. These NSCyanine dyes with a low pKa (<4.0) only show weak fluorescence at lysosome pH (key1), however, the fluorescence can be completely switched on and significantly enhanced by intracellular viscosity (key2) in disease tissues, exhibiting high target-to-liver ratios up to 19.5/1. Notably, high-contrast phototheranostics in extrahepatic diseases are achieved, including intestinal metastasis-imaging, acute gastritis-imaging, bacteria infected wound healing, and tumor ablation via targeted combined photothermal therapy and chemotherapy.

H2O2-Activated NIR-II Fluorescent Probe with a Large Stokes Shift for High-Contrast Imaging in Drug-Induced Liver Injury Mice.

Drug-induced liver injury (DILI) is the most common clinical adverse drug reaction, which is closely associated with the oxidative stress caused by overproduced reactive oxygen species. Hepatic H2O2, as an important biomarker of DILI, plays a crucial role in the progression of DILI. However, there remains a challenge to develop H2O2-activatable second near-infrared (NIR-II, 1000-1700 nm) small molecular probes with both a large Stokes shift and a long emission wavelength beyond 950 nm. Herein, we developed an activatable NIR-II fluorescent probe (IR-990) with an acceptor-π-acceptor (A-π-A) skeleton for real-time detection of H2O2 in vivo. In the presence of H2O2, nonfluorescent probe IR-990 was successfully unlocked by generating a donor-π-acceptor (D-π-A) structure and switched on intense NIR-II fluorescence, exhibiting a peak emission wavelength at 990 nm and a large Stokes shift of 200 nm. Moreover, it was able to detect H2O2 with high sensitivity and selectivity in vitro (LOD = 0.59 µM) and monitor the behavior of endogenous H2O2 in the HepG2 cell model of DILI for the first time. Notably, probe IR-990 was successfully applied in real-time imaging of endogenous H2O2 generation in the DILI mouse model, showing a high signal-to-background ratio of 11.3/1. We envision that IR-990 holds great potential as a powerful diagnosis tool for real-time visualization of H2O2 in vivo and revealing the mechanism of DILI in the future.

Bichromatic Imaging with Hemicyanine Fluorophores Enables Simultaneous Visualization of Non-alcoholic Fatty Liver Disease and Metastatic Intestinal Cancer.

Simultaneous detection of different diseases via a single fluorophore is challenging. We herein report a bichromatic fluorophore named Cy-914 for the simultaneous diagnosis of non-alcoholic fatty liver disease (NAFLD) and metastatic intestinal cancer by leveraging its NIR-I/NIR-II dual-color imaging capability. Cy-914 with a pKa of 6.98 exhibits high sensitivity to pH and viscosity, showing turn-on NIR-I fluorescence at 795 nm in an acidic tumor microenvironment, meanwhile displaying intense NIR-II fluorescence at 914/1030 nm under neutral to slightly basic viscous conditions. Notably, Cy-914 could sensitively and noninvasively monitor viscosity variations in the progression of NAFLD. More importantly, it was able to simultaneously visualize NAFLD (ex/em = 808/1000-1700 nm) and intestinal metastases (ex/em = 570/810-875 nm) in two independent channels without spectral cross interference after topical spraying, further improving fluorescence-guided surgery of tiny metastases less than 3 mm. This strategy may provide an understanding for developing multi-color fluorophores for multi-disease diagnosis.

Three Birds with One Stone: Acceptor Engineering of Hemicyanine Dye with NIR-II Emission for Synergistic Photodynamic and Photothermal Anticancer Therapy.

It is challenging to develop a near-infrared (NIR) small molecular photosensitizer for synergistic phototherapy in deep tissues. Herein, first, a heavy-atom-free NIR hemicyanine photosensitizer (BHcy) for 808 nm light-mediated synergistic photodynamic therapy/photothermal therapy (PDT/PTT) anticancer therapy by leveraging the acceptor engineering strategy is reported. This strategy endows BHcy with a more planar and larger π-conjugated structure, resulting in long NIR absorption/emission at 770/915-1200 nm as well as enhanced singlet oxygen (1 O2 ) generation ability and photothermal effect, which is ascribed to the reduced energy levels of excited singlet/triplet states and the promoted intersystem crossing process. Notably, BHcy-based nanoparticles (BHcy-NPs) exhibit efficient 1 O2 yield (12.9%) and high photothermal conversion efficiency (55.1%). More importantly, BHcy-NPs are able to significantly kill cancer cells by destroying main organelles and inhibit tumor growth in vivo after a single irradiation. Overall, this study provides a strategy to design new heavy-atom-free PDT/PTT agents for potential clinical applications.

Plants Mitigate Nitrous Oxide Emissions from Antibiotic-Contaminated Agricultural Soils.

Vegetable production systems are hotspots of nitrous oxide (N2O) emissions and antibiotic pollution. However, little is known about the interconnections among N2O emissions, vegetable growth, and antibiotic contamination. To understand how plants regulate N2O emissions from enrofloxacin (ENR)-contaminated soils, in situ N2O emissions were measured in pot experiments with cherry radish and pakchoi. Gross N2O production and consumption processes were discriminated based on an acetylene inhibition experiment. Results indicated that vegetable growth decreased the cumulative N2O flux from 0.71 to -0.29 kg ha-1 and mitigated the ENR-induced increase in N2O emissions. Radish displayed better mitigation of N2O emissions than pakchoi. By combining the analysis of N2O flux with soil physicochemical and microbiological properties, we demonstrated that growing vegetables could either promote gross N2O consumption or decrease gross N2O production, primarily by interacting with soil nitrate, clade II nosZ (nosZII)-carrying bacteria, and Deinococcus-Thermus. ENR inhibited N2O consumption more than N2O production, with the nosZII-carrying bacteria, represented by Gemmatimonadetes, as the main inhibition target. However, increasing nosZII-carrying bacteria by growing radish offsets the inhibitory effect of ENR. These findings provide new insights into N2O emissions and antibiotic pollution in vegetable-soil ecosystems and broaden the options for mitigating N2O emissions.

Clinical findings of Talaromyces marneffei infection among patients with anti-interferon-γ immunodeficiency: a prospective cohort study.

BACKGROUND: Talaromyces marneffei (T. marneffei) infection has been associated with adult-onset immunodeficiency due to anti-IFN-γ autoantibodies. We aimed to investigate the clinical features of non-HIV-infected patients with T. marneffei infection in southern China. METHODS: Between January 2018 and September 2020, we enrolled patients with T. marneffei infection who were HIV-negative (group TM, n = 42), including anti-IFN-γ autoantibody-positive (group TMP, n = 22) and anti-IFN-γ autoantibody-negative (group TMN, n = 20) patients and healthy controls (group HC, n = 40). Anti-IFN-γ autoantibodies were detected by ELISA. Clinical characteristics and clinical laboratory parameters were recorded. RESULTS: Compared with anti-IFN-γ autoantibody-negative patients with T. marneffei infection, anti-IFN-γ autoantibody-positive patients did not have underlying respiratory disease; more frequently exhibited dissemination of systemic infections with severe pleural effusion; had higher WBC counts, C-reactive protein levels, erythrocyte sedimentation rates, and neutrophil and CD8+ T cell counts; had lower hemoglobin levels; and were more likely to have other intracellular pathogen infections. Most of these patients had poor outcomes despite standardized antimicrobial therapy. CONCLUSION: T. marneffei-infected patients with higher anti-IFN-γ autoantibody titers have more severe disease and complex clinical conditions.

A New Classifier Based on Laboratory Indicators for Early Diagnosis and Prognosis Prediction of Ewing's Sarcoma.

BACKGROUND: Ewing's sarcoma (ES) is a prevalent bone malignancy. It is critical to explore new diagnostic and prognostic indicators because of the rapid progression of ES and the low survival rate of metastatic ES patients. However, few parameters of clinical significance have been found. The aim of this study was to establish a new classifier with clinical laboratory data to help ES detection and prognosis prediction. METHODS: A total of 135 ES patients, 150 healthy individuals, and 228 patients with primary benign bone lesions were included. Logistic regression on clinical laboratory indicators was conducted to establish the classifier, and then the classifier was assessed by drawing the receiver operating characteristic (ROC) curves. Patient survival was evaluated using the Kaplan-Meier method. RESULTS: We established the diagnostic classifier, called Ces, with clinical laboratory indicators to distinguish ES from healthy individuals. Ces showed great diagnostic performance in the test cohort (area under the receiver operating characteristic curve (AUC) 0.95) and could identify early-stage (AUC 0.93) and small-size (AUC 0.95) ES effectively. In addition, the classifier had good ability to differentiate ES from primary benign bone lesions (AUC 0.77 for Ces, AUC 0.83 for Ces + age). Furthermore, Ces was associated with tumor metastasis and event-free survival (EFS) of ES patients and showed better performance than lactate dehydrogenase (LDH) in prognosis prediction. CONCLUSIONS: Our study indicates that Ces has the potential to be a non-invasive biomarker for ES diagnosis and prognosis.

Novel classifiers with clinical laboratory parameters for early detection of osteosarcoma.

BACKGROUND: Osteosarcoma (OS) is one of the most common malignant bone tumors. It is essential to explore early diagnostic indicators with high sensitivity and specificity due to the rapid progression and metastasis of OS and the poor survival of metastatic OS patients. However, a few indicators of diagnostic significance have been described. METHODS: A total of 458 OS patients, 312 healthy individuals, and 228 patients with primary benign bone lesions were included. Logistic regression was performed on 46 clinical laboratory parameters to establish the diagnostic classifiers, which were evaluated by analysis of the receiver operating characteristic (ROC) curves. RESULTS: We established three diagnostic classifiers, called Cos for all ages, Clos for low ages, and Chos for high ages, with clinical laboratory parameters to distinguish OS from healthy individuals. All classifiers showed better diagnostic performances than alkaline phosphatase (ALP) in the independent validation cohort. In addition, these classifiers had better ability than ALP to discriminate OS from primary benign bone lesions. Furthermore, Cos , Clos, and Chos had larger AUC than ALP to identify small-size and early-stage OS and could also detect ALP-negative OS effectively. CONCLUSION: Our study suggests the potential of Cos , Clos , and Chos as non-invasive biomarkers for early OS.

Fungal community composition change and heavy metal accumulation in response to the long-term application of anaerobically digested slurry in a paddy soil.

Anaerobically digested slurry (ADS) has been widely used as a liquid fertilizer in agroecosystems. However, there is scant information on the effects of successive ADS applications on heavy metals (HMs) accumulation and fungal community composition in paddy soils. In this study, we conducted a field experiment over 10 years to assess the changes in soil HMs and fungal community composition under the long-term application of ADS in a paddy field. The four treatments were (1) no fertilizer (CK); (2) mineral fertilizer and 270 kg N ha-1 from urea (MF); (3) 270 kg N ha-1 from ADS (ADS1); and (4) 540 kg N ha-1 from ADS (ADS2). The results revealed that ADS application improved paddy soil fertility compared to that under the MF treatment by increasing soil organic C (SOC), total N (TN) and available potassium (AK). Long-term application of ADS significantly increased soil total and available Zn (TZn and AZn) concentrations as compared to those under the MF treatment. However, there were no significant differences in the total and available Cu concentrations or the total Pb concentration between the ADS and MF treatments. Sequence analysis showed that application of ADS increased the fungal richness indexes (Chao1 and ACE) compared to MF treatment. Principal coordinate analysis (PCoA) showed that the soil fungal community compositions were significantly separated by high levels of ADS application. Long-term application of ADS increased the relative abundance of classes Sordariomycetes, Dothideomycetes and Agaricomycetes by 20.8-29.0%, 107.3-141.4% and 289.5-387.5%, respectively, but decreased that of Pezizomycetes by 14.0-33.0% compared to that under the MF treatment. At the genus level, compared to those under the MF treatment, the relative abundances of Pyrenochaetopsis and Myrothecium were significantly increased by the application of ADS, but those of Mrakia and Tetracladium were significantly decreased. Redundancy analysis (RDA) revealed that SOC, AZn and AP were the three most important factors affecting the fungal community composition of the paddy soil. Our findings suggested that fungal community composition could be affected by changes in the chemical properties and heavy metal contents of paddy soil under high application of ADS in the long term.

Multifunctional oligomer immobilized on quartz crystal microbalance: a facile and stabilized molecular imprinting strategy for glycoprotein detection.

Glycoprotein detection holds great potential for early diagnosis of diverse diseases. For this purpose, the combination of quartz crystal microbalance (QCM) sensor and molecular imprinting has attracted increasing attention. Nonetheless, the recently common imprinted films fabricated on QCM electrode are thick and rigid, lacking flexibility in aqueous phase. Alternatively, small molecules immobilized on the electrode to construct molecular scale film could address this problem, while stabilization of the imprinted sites remains challenging. Herein, a co-assembly complex was obtained by the mixture of template and multifunctional oligomer, which was then immobilized on the amino-modified transducer surface through epoxy-amino reaction to form a protein-imprinted film. Afterward, the remaining epoxy groups in oligomer chains were cross-linked to conserve and stabilize the orientation of imprinted sites after template elution. Template rebinding tests show that cross-linked film has much higher imprinting factors than that of the non-cross-linked counterpart. Furthermore, control proteins that are distinct in properties and structures were employed to demonstrate the selectivity of this approach, and the imprinted assay reveals high affinity and specificity towards template protein. Graphical Abstract.

How robot-assisted gait training affects gait ability, balance and kinematic parameters after stroke: a systematic review and meta-analysis.

INTRODUCTION: Gait ability is often cited by stroke survivors. Robot-assisted gait training (RAGT) can help stroke patients with lower limb motor impairment regain motor coordination. EVIDENCE ACQUISITION: PubMed, Cochrane Library, Embase were systematically searched until September 2023, to identify randomized controlled trials presenting: stroke survivors as participants; RAGT as intervention; conventional rehabilitation as a comparator; gait assessment, through scales or quantitative parameters, as outcome measures. EVIDENCE SYNTHESIS: Twenty-seven publications involving 1167 patients met the inclusion criteria. Meta-analysis showed no significant differences in speed, cadence, spatial symmetry, and changes in joint mobility angles between the RAGT group and the control group. In addition, RAGT was associated with changes in affected side step length (SMD=0.02, 95% CI: 0.01, 0.03; P<0.0001), temporal symmetry (SMD=-0.38, 95% CI: -0.6, -0.16; P=0.0006], Six-Minute Walk Test (SMD=25.14, 95% CI: 10.19, 40.09; P=0.0010] and Functional Ambulation Categories (SMD=0.32, 95% CI: 0.01, 0.63; P=0.04). According to the PEDro scale, 19 (70.4%) studies were of high quality and eight were of moderate quality (29.6%). CONCLUSIONS: Taken together, the review synthesis showed that RAGT might have a potential role in the recovery of walking dysfunction after stroke. However, its superiority over conventional rehabilitation requires further research. Additionally, it may provide unexpected benefits that the effects of RAGT with different types or treatment protocols were further compared.