Genomic epidemiology reveals early transmission of SARS-CoV-2 and mutational dynamics in Nanning, China.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are a fatal pathogen resulting in substantial morbidity and mortality, and posing a great threat to human health with epidemics and pandemics. METHODS: Next-generation sequencing (NGS) was performed to investigate the SARS-CoV-2 genomic characterization. Phylogenetic analysis of SARS-CoV-2 genomes was used to probe the evolutionary. Homology protein structure modelling was done to explore potential effect of the mutations. RESULTS: The eighty genome sequences of SARS-CoV-2 obtained from the thirty-nine patients with COVID-19. A novel variant with mutation H625R concomitant with S50L in spike glycoprotein had been identified. Phylogenetic analysis revealed that SARS-CoV-2 variants belong to several distinct lineages. Homology modelling indicated that variant with mutation H625R and S50L increases flexibility of S1 subunit. CONCLUSIONS: SARS-CoV-2 genomes are constantly evolving by accumulation of point mutations. The amino acid H625R in combination with S50L may have a significant impact on the interaction between spike glycoprotein and ACE2.

Whale Optimization Algorithm with a Hybrid Relation Vector Machine: A Highly Robust Respiratory Rate Prediction Model Using Photoplethysmography Signals.

Due to the simplicity and convenience of PPG signal acquisition, the detection of the respiration rate based on the PPG signal is more suitable for dynamic monitoring than the impedance spirometry method, but it is challenging to achieve accurate predictions from low-signal-quality PPG signals, especially in intensive-care patients with weak PPG signals. The goal of this study was to construct a simple model for respiration rate estimation based on PPG signals using a machine-learning approach fusing signal quality metrics to improve the accuracy of estimation despite the low-signal-quality PPG signals. In this study, we propose a method based on the whale optimization algorithm (WOA) with a hybrid relation vector machine (HRVM) to construct a highly robust model considering signal quality factors to estimate RR from PPG signals in real time. To detect the performance of the proposed model, we simultaneously recorded PPG signals and impedance respiratory rates obtained from the BIDMC dataset. The results of the respiration rate prediction model proposed in this study showed that the MAE and RMSE were 0.71 and 0.99 breaths/min, respectively, in the training set, and 1.24 and 1.79 breaths/min, respectively, in the test set. Compared without taking signal quality factors into account, MAE and RMSE are reduced by 1.28 and 1.67 breaths/min, respectively, in the training set, and reduced by 0.62 and 0.65 breaths/min in the test set. Even in the nonnormal breathing range below 12 bpm and above 24 bpm, the MAE reached 2.68 and 4.28 breaths/min, respectively, and the RMSE reached 3.52 and 5.01 breaths/min, respectively. The results show that the model that considers the PPG signal quality and respiratory quality proposed in this study has obvious advantages and application potential in predicting the respiration rate to cope with the problem of low signal quality.

Viral Dynamic Surveillance in COVID-19 Patients: A Cohort Study.

Background: Coronavirus disease 2019 (COVID-19) is a potentially fatal pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), especially those of novel SARS-CoV-2 variants and infection has affected over 700 million people globally. Methods: This retrospective, descriptive study included 118 patients admitted with SARS-CoV-2 infection as confirmed by real-time reverse transcription polymerase chain reaction. Results: The median duration of detectable SARS-CoV-2 infection in patients with high ALT, AST, and PLT/LYMPH, or low CD4+, CD8+, and PLT/MONO was considerably longer. In the risk factor model, multivariate analysis was performed for the estimation of ALT (HR, 0.54; 95% CI, 0.36-0.81), AST (HR, 0.56; 95% CI, 0.34-0.93), CD4+ (HR,0.77; 95% CI, 0.48-1.24), CD8+ (HR,0.64; 95% CI, 0.37-1.11), PLT/LYMPH (HR, 1.16; 95% CI, 0.76-1.77), and PLT/MONO (HR, 0.64; 95% CI, 0.43-0.94). Conclusions: The longer viral RNA duration was associated with a higher International Prognostic Index score (p = 0.0013), demonstrating for the first time that multivariate features of the bioindicators closely associated with SARS-CoV-2-infected patients clear the virus.

Painless and sensitive pepsinogen I detection: an electrochemical immunosensor based on rhombic dodecahedral Cu3Pt and MoS2 NFs.

Gastric cancer (GC) is a common malignant tumour of the digestive tract with a high mortality rate worldwide. However, many patients delay treatment due to the avoidance of the costly and painful procedure of gastroscopy. Therefore, an early convenient screening method is essential to improve the survival rate of GC patients. To address this issue, we constructed an electrochemical immunosensor supported by rhombohedral Cu3Pt and MoS2 nanoflowers (MoS2 NFs) for rapid, painless and quantitative detection of the GC biomarker in vitro. Here, pepsinogen I was employed as a model protein biomarker to analyse the performance of the immunosensor. The rhombohedral dodecahedral Cu3Pt nanoparticles decorated with MoS2-NFs were further functionalized; this allowed the constructed sensor to possess more nano- or micro-structures, thereby improving the detection sensitivity. In specific applications, the corresponding bioactive molecules can be flexibly captured. Under optimal conditions, the immunoassay showed a wide linear range from 500 pg mL-1 to 400 ng mL-1 and a low detection limit of 167 pg mL-1 (S/N = 3). This covers the critical value of 70 ng mL-1, and the results obtained from the analysis of human serum samples were on par with those from the enzyme immunoassay, suggesting significant potential for this new method in daily diagnosis.

A rapid lateral flow immunoassay strip for detection of SARS-CoV-2 antigen using latex microspheres.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly infectious and concealed virus that causes pneumonia, severe acute respiratory syndrome, and even death. Although the epidemic has been controlled since the development of vaccines and quarantine measures, many people are still infected, particularly in third-world countries. Several methods have been developed for detection of SARS-CoV-2, but owing to its price and efficiency, the immune strip could be a better method for the third-world countries. METHODS: In this study, two antibodies were linked to latex microspheres, using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, as the bridge to decrease the cost further and improve the detection performance. The specificity of the lateral flow immunoassay strip (LFIA) was tested by several common viruses and respiratory bacterial infections. Besides, the reproducibility and stability of the LFIAs were tested on the same batch of test strips. Under optimal conditions, the sensitivity of LFIA was determined by testing different dilutions of the positive specimens. RESULTS: The proposed LFIAs were highly specific, and the limit of detection was as low as 25 ng/mL for SARS-CoV-2 antigens. The clinical applicability was evaluated with 659 samples (230 positive and 429 negative samples) by using both LFIA and rRT-PCR. Youden's index (J) was used to assess the performance of these diagnostic tests. The sensitivity and specificity were 98.22% and 97.93%, respectively, and J is 0.9615. The sensitivity and specificity were 98.22% and 97.93%, respectively, and J is 0.9615. In addition, the consistency of our proposed LFIA was analyzed using Cohen's kappa coefficient (κ = 0.9620). CONCLUSION: We found disease stage, age, gender, and clinical manifestations have only a slight influence on the diagnosis. Therefore, the lateral flow immunoassay SARS-CoV-2 antigen test strip is suitable for point-of-care detection and provides a great application for SARS-CoV-2 epidemic control in the third-world countries.

Electrochemical immunosensor based on binary nanoparticles decorated rGO-TEPA as magnetic capture and Au@PtNPs as probe for CEA detection.

Using gold and magnetic nanoparticles co-decorated reduced graphene oxide-tetraethylenepentamine (rGO-TEPA/Au-MNPs) as the magnetic platform for capturing the primary antibody (Ab1), separation and preconcentration of immunocomplex, a novel homogeneous electrochemical immunosensor was successfully developed. The newly prepared magnetic rGO-TEPA/Au-MNPs, compared with MNPs, exhibited better stability and enhanced electrical conductivity attributed to rGO-TEPA, and showed higher biorecognition efficiency due to AuNPs. In addition, Au@PtNPs were prepared and modified with secondary antibody (Ab2) as an efficient signal probe for signal readout. Using carcinoembryonic antigen (CEA) as a model analyte, the prepared immunosensor demonstrated satisfactory properties like high stability, good repeatability and selectivity, wide linear range (5.0 pg mL-1~200.0 ng mL-1) as well as low detection limit (1.42 pg mL-1). The homogenous electrochemical immunosensor was applied to the detection of CEA in human serum and was found to exhibit good correlation with the reference method. Thus, the proposed rGO-TEPA/Au-MNPs-based homogenous immunoassay platform might open up a new way for biomarker diagnosis. Graphical Abstract.

A Label-Free Immunosensor Based on Graphene Oxide/Fe3O4/Prussian Blue Nanocomposites for the Electrochemical Determination of HBsAg.

In this article, a highly sensitive label-free immunosensor based on a graphene oxide (GO)/Fe3O4/Prussian blue (PB) nanocomposite modified electrode was developed for the determination of human hepatitis B surface antigen (HBsAg). In this electrochemical immunoassay system, PB was used as a redox probe, while GO/Fe3O4/PB nanocomposites and AuNPs were prepared and coated on screen-printed electrodes to enhance the detection sensitivity and to immobilize the hepatitis B surface antibody (HBsAb). The immunosensor was fabricated based on the principle that the decrease in peak currents of PB is proportional to the concentration of HBsAg captured on the modified immunosensor. The experimental results revealed that the immunosensor exhibited a sensitive response to HBsAg in the range of 0.5 pg·mL-1 to 200 ng·mL-1, and with a low detection limit of 0.166 pg·mL-1 (S/N = 3). Furthermore, the proposed immunosensor was used to detect several clinical serum samples with acceptable results, and it also showed good reproducibility, selectivity and stability, which may have a promising potential application in clinical immunoassays.

A novel 3D paper-based microfluidic electrochemical glucose biosensor based on rGO-TEPA/PB sensitive film.

A novel 3D paper-based microfluidic screen-printed electrode (SPE) composed of two layers was constructed by photolithography and screen-printing technology. Aldehyde functionalized hydrophilic zone of the counter and reference electrodes layer was prepared for glucose oxidase immobilization. Highly conductive prussian blue deposited reduced graphene oxide-tetraethylene pentamine (rGO-TEPA/PB) modified paper working electrode layer can be used as an electrochemical sensitive membrane for quantitative detection of hydrogen peroxide (H2O2), which was the enzyme-catalyzed reaction product. Therefore, this 3D paper-based microfluidic electrochemical biosensor can be used for quantitative detection of glucose. Under optimum conditions, the proposed biosensor can be used for quantitative determination of glucose over a wide linear range of 0.1 mM-25 mM with detection limit of 25 µM. Finally, the 3D paper-based microfluidic electrochemical biosensor was applied to determine glucose in human sweat and blood, and the obtained results were in good consistency with values measured by Roche's blood glucose meter. In addition, the proposed 3D paper-based electrochemical device showed good repeatability, stability, and anti-interference, which would be of great potential to monitor glucose in complex biological fluids.

Disposable Amperometric Immunosensor for Hepatitis B Antigen Detection Based on Multiwalled Carbon Nanotubes and Ferrocene Decorated Screen Printed Electrode.

In this work, we report an amperometric immunosensor for detecting Hepatitis B surface antigen based on ChitosanFerrocene-Ammoniated multiwalled carbon nanotubes (CS-Fc-AMWNTs). The CS-Fc-AMWNTs nanocomposites were produced by Schiff base reaction, providing not only large specific surface area, but also favorable conductivity and outstanding biocompatibility. HBsAb was modified on electrode surface by glutaraldehyde cross-linking. The immunocomplex was produced after incubation with HBsAg, which hindered the electron transfer and concentration of antibody was observed to change in the current. The immunosensor showed great linear range from 1-250 ng/mL. Results from this study show that the immunosensor has good reliability and high sensitivity for detection of HBsAg in certain clinical application value.

Hollow Fe3O4/Graphene Oxide Nanocomposites as Novel Rapamycin Carrier: Formulation Optimization and In Vitro Characterization.

In the present research an attempt was made to develop and optimize rapamycin (Rapa) loaded hollow magnetic Fe3O4/graphene oxide (Fe3O4/GO) nanocomposites using solvent evaporation technique and response surface methodology (RSM). A Box-Behnken design (BBD) with a three-level, three-factor was used to determine preparation parameters that would achieve the highest encapsulation efficiency (EE) and drug loading capacity (DLC). At optimum conditions such as mass ratio of Rapa to Fe3O4/GO (Rapa: Fe3O4/GO) 0.45, oscillation rate 144.0, and volume ratio of water to solvent (W/S) 4.2, the EE and DLC of experimentally prepared particles reached 84.92 ± 5.50% and 28.68 ± 2.54% respectively. The morphological assessment results showed that hollow Fe3O4 nano-aggregates were evenly and tightly dispersed on the layer of GO membrane. After the addition of GO and encapsulation of Rapa, the Fe3O4/GO nanocomposites and Rapa loaded magnetic Fe3O4/GO (Fe3O4/GO-Rapa) nanocomposites showed saturation magnetization of 57.77 and 40.71 emu/g respectively. The drug releasing experiment indicated a slightly acidic atmosphere, which was suitable for Rapa releasing from the obtained Fe3O4/GO nanocomposites. The cell counting Kit-8 (CCK-8) assays demonstrated the hollow Fe3O4/GO nanocomposites could effectively improve the efficacy of Rapa in killing HepG2 cells, which displayed a concentration-dependent manner. All these results suggested the prepared Fe3O4/GO nanocomposites have a potential application in drug delivery system.

Paper-based microfluidic devices for electrochemical immunofiltration analysis of human chorionic gonadotropin.

An electrochemical immunofiltration analysis was introduced into microfluidic paper-based analytical devices (µPADs) for the first time, which was based on photolithography and screen-printing technology. The hydrophilic test zones of the aldehyde-functionalized screen-printed electrodes (SPEs) were biofunctionalized with capture antibodies (Ab1). A sensitive immune detection method was developed by using primary signal antibody functionalized gold nanoparticles (GNPs/Ab2) and alkaline phosphatase conjugated secondary antibody (ALP-IgG). Differential pulse voltammetry (DPV) was performed to detect the electrochemical response. The microfluidic paper-based electrochemical immunosensor (µ-PEI) was optimized and characterized for the detection of human chorionic gonadotropin (HCG), a model analyte, in a linear range from 1.0mIUmL-1 to 100.0 IU mL-1 with a detection limit of 0.36mIUmL-1. Additionally, the proposed µ-PEI was used to test HCG in real human serum and obtained satisfactory results. The disposable, efficient, sensitive and low-cost µ-PEI has exhibited great potential for the development of point-of-care testing (POCT) devices that can be applicated in healthcare monitoring.

Chelerythrine and Fe3O4 Loaded Multi-Walled Carbon Nanotubes for Targeted Cancer Therapy.

The work focused on manufacturing improved drug loaded multifunctional magnetic nanoparticles that can overcome the relative non-specificity and potential side-effects of some chemotherapeutic drugs to healthy tissues. A new drug delivery system, Chelerythrine (CHE) and Fe3O4 loaded multi-walled carbon nanotubes (Fe3O4/MWNTs-CHE nanocomposites) that can target hepatocytes when treating malignant tumors, was prepared through a simple adsorption method. The formulation and structure of the Fe3O4/MWNTs-CHE nanocomposites were characterized by vibrating sample magnetometer (VSM), Fourier Transform infrared spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The cytotoxicity and anti-proliferation effect from the prepared nanocomposites were in vitro tested on human hepatocarcinoma HepG2 and normal liver LO2 cell lines. The results showed the saturated magnetization of Fe3O4/MWNTs-CHE nanocomposites could reach to 45.4O3 emu/g, and the in vitro CHE release behavior exhibited a biphasic release pattern. Moreover, the in vitro cytotoxicity studies revealed that the Fe3O4/MWNTs-CHE nanocomposites showed an efficient inhibition rate to HepG2 cell line and exhibited a lower cytotoxicity to LO2 cell line in comparison to the native CHE. Therefore, the multifunctional Fe3O4/MWNTs-CHE nanocomposites should be a useful and promising candidate for treatment of malignant tumors.

Rapamycin loaded magnetic Fe3O4/carboxymethylchitosan nanoparticles as tumor-targeted drug delivery system: Synthesis and in vitro characterization.

A novel tumor-targeted drug delivery system (Fe3O4/CMCS-Rapa NPs) was prepared using magnetic Fe3O4/carboxymethylchitosan nanoparticles (Fe3O4/CMCS NPs) as carrier and rapamycin (Rapa) as the model anti-tumor drug. The morphology, composition, and properties of the Fe3O4/CMCS-Rapa NPs were characterized by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscope (TEM), X-ray diffraction (XRD), thermal analysis (TG/DSC), vibration sample magnetometer (VSM), and drug release kinetics, cytotoxicity, cellular uptake, apoptosis studies in vitro. The results showed that the synthesized Fe3O4/CMCS-Rapa NPs were spherical in shape with an average size of 30±2 nm, the saturated magnetization reached 67.1 emu/g, and the loading efficiency of Rapa was approximately 6.32±0.34%. In addition, the in vitro drug release behavior displayed that the Fe3O4/CMCS NPs exhibited a biphasic drug release pattern with initial burst release and consequently sustained release. Furthermore, the Fe3O4/CMCS-Rapa NPs showed lower cytotoxicity to liver cell line (LO2) and comparatively higher cytotoxicity to human hepatocarcinoma cell line (HepG2) than native Rapa. Fe3O4/CMCS-Rapa NPs could enhance cellular uptake and reduce Rapa drug damage to the normal cells so as to improve the curative effect of drug to tumor cells. All these results demonstrated that the Fe3O4/CMCS-Rapa NPs may be useful as a promising candidate for targeted cancer diagnostic and therapy.