Preparation of PEI-modified nanoparticles by dopamine self-polymerization for efficient DNA delivery.

Achieving efficient and safe gene delivery is of great significance to promote the development of gene therapy. In this work, a polydopamine (PDA) layer was coated on the surface of Fe3 O4 nanoparticles (NPs) by dopamine (DA) self-polymerization, and then magnetic Fe3 O4 NPs were prepared by the Michael addition between amino groups in polyethyleneimine (PEI) and PDA. The prepared Fe3 O4 NPs (named Fe3 O4 @PDA@PEI) were characterized by Fourier transform infrared (FTIR), atomic force microscopy (AFM), and scanning electron microscopy (SEM). As an efficient and safe gene carrier, the potential of Fe3 O4 @PDA@PEI was evaluated by agarose gel electrophoresis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, fluorescence microscopy, and flow cytometry. The results show that the Fe3 O4 @PDA@PEI NPs are stable hydrophilic NPs with a particle size of 50-150 nm. It can efficiently condense DNA at low N/P ratios and protect it from nuclease degradation. In addition, the Fe3 O4 @PDA@PEI NPs have higher safety than PEI. Further, the Fe3 O4 @PDA@PEI/DNA polyplexes could be effectively absorbed by cells and successfully transfected and exhibit higher cellular uptake and gene transfection efficiency than PEI/DNA polyplexes. The findings indicate that the Fe3 O4 @PDA@PEI NPs have the potential to be developed into a novel gene vector.

The structural and molecular mechanisms of type II PETases: a mini review.

The advent of plastics has led to significant advances for humans, although the accompanying pollution has also been a source of concern for countries globally. Consequently, a biological method to effectively degrade polyethylene terephthalate (PET) has been an area of significant scientific interest. Following the report of the highly efficient PET hydrolase from the bacterium Ideonella sakaiensis strain 201-F6 (i.e., IsPETase) in 2016, its structure has been extensively studied, showing that it belongs to the type II PETase group. Unlike type I PETases that include most known cutinases, structural investigations of type II PETases have only been conducted since 2017. Type II PETases are further divided into type IIa and IIb enzymes. Moreover, even less research has been conducted on type IIa plastic-degrading enzymes. Here, we present a review of recent studies of the structure and mechanism of type II PETases, using the known structure of the type IIa PETase PE-H from the marine bacterium Pseudomonas aestusnigri in addition to the type IIb enzyme IsPETase as representatives. These studies have provided new insights into the structural features of type II PETases that exhibit PET catalytic activity. In addition, recent studies investigating the rational design of IsPETases are reviewed and summarized alongside a discussion of controversies surrounding PETase investigations.

Deep learning model for multi-classification of infectious diseases from unstructured electronic medical records.

PURPOSE: Predictively diagnosing infectious diseases helps in providing better treatment and enhances the prevention and control of such diseases. This study uses actual data from a hospital. A multiple infectious disease diagnostic model (MIDDM) is designed for conducting multi-classification of infectious diseases so as to assist in clinical infectious-disease decision-making. METHODS: Based on actual hospital medical records of infectious diseases from December 2012 to December 2020, a deep learning model for multi-classification research on infectious diseases is constructed. The data includes 20,620 cases covering seven types of infectious diseases, including outpatients and inpatients, of which training data accounted for 80%, i.e., 16,496 cases, and test data accounted for 20%, i.e., 4124 cases. Through the auto-encoder, data normalization and sparse data densification processing are carried out to improve the model training effect. A residual network and attention mechanism are introduced into the MIDDM model to improve the performance of the model. RESULT: MIDDM achieved improved prediction results in diagnosing seven kinds of infectious diseases. In the case of similar disease diagnosis characteristics and similar interference factors, the prediction accuracy of disease classification with more sample data is significantly higher than the prediction accuracy of disease classification with fewer sample data. For instance, the training data for viral hepatitis, influenza, and hand foot and mouth disease were 2954, 3924, and 3015 respectively and the corresponding test accuracy rates were 99.86%, 98.47%, and 97.31%. There is less training data for syphilis, infectious diarrhea, and measles, i.e., 1208, 575, and 190 respectively and the corresponding test accuracy rates were noticeably lower, i.e., 83.03%, 87.30%, and42.11%. We also compared the MIDDM model with the models used in other studies. Using the same input data, taking viral hepatitis as an example, the accuracy of MIDDM is 99.44%, which is significantly higher than that of XGBoost (96.19%), Decision tree (90.13%), Bayesian method (85.19%), and logistic regression (91.26%). Other diseases were also significantly better predicted by MIDDM than by these three models. CONCLUSION: The application of the MIDDM model to multi-class diagnosis and prediction of infectious diseases can improve the accuracy of infectious-disease diagnosis. However, these results need to be further confirmed via clinical randomized controlled trials.

Glycerol Production from Undetoxified Lignocellulose Hydrolysate by a Multiresistant Engineered Candida glycerinogenes.

Glycerol is an important platform compound with multidisciplinary applications, and glycerol production using low-cost sugar cane bagasse hydrolysate is promising. Candida glycerinogenes, an industrial yeast strain known for its high glycerol production capability, has been found to thrive in bagasse hydrolysate obtained through a simple treatment without detoxification. The engineered C. glycerinogenes exhibited significant resistance to furfural, acetic acid, and 3,4-dimethylbenzaldehyde within undetoxified hydrolysates. To further enhance glycerol production, genetic modifications were made to Candida glycerinogenes to enhance the utilization of xylose. Fermentation of undetoxified bagasse hydrolysate by CgS45 resulted in a glycerol titer of 40.3 g/L and a yield of 40.4%. This process required only 1 kg of bagasse to produce 93.5 g of glycerol. This is the first report of glycerol production using lignocellulose, which presents a new way for environmentally friendly industrial production of glycerol.

PEI functionalized cell membrane for tumor targeted and glutathione responsive gene delivery.

Polyethylenimine (PEI) is a broadly exploited cationic polymer due to its remarkable gene-loading capacity. However, the high cytotoxicity caused by its high surface charge density has been reported in many cell lines, limiting its application significantly. In this study, two different molecular weights of PEI (PEI10k and PEI25k) were crosslinked with red blood cell membranes (RBCm) via disulfide bonds to form PEI derivatives (RMPs) with lower charge density. Furthermore, the targeting molecule folic acid (FA) molecules were further grafted onto the polymers to obtain FA-modified PEI-RBCm copolymers (FA-RMP25k) with tumor cell targeting and glutathione response. In vitro experiments showed that the FA-RMP25k/DNA complex had satisfactory uptake efficiency in both HeLa and 293T cells, and did not cause significant cytotoxicity. Furthermore, the uptake and transfection efficiency of the FA-RMP25k/DNA complex was significantly higher than that of the PEI25k/DNA complex, indicating that FA grafting can increase transfection efficiency by 15 %. These results suggest that FA-RMP25k may be a promising non-viral gene vector with potential applications in gene therapy.

The damages of high intensity focused ultrasound to transplanted hydatid cysts in abdominal cavities of rabbits with aids of ultrasound contrast agent and superabsorbent polymer.

The present study investigates the damages of high intensity focused ultrasound (HIFU) to transplanted hydatid cysts in abdominal cavities of rabbits with aids of ultrasound contrast agent (UCA) and superabsorbent polymer (SAP) alone or in combination. A rabbit model with transplanted hydatid cyst was established by implanting hydatid cyst isolated from infected sheep liver, and HIFU was used to ablate the transplanted cysts with the aid of UCA and SAP alone or in combination. The hydatid cyst with thin wall, good elasticity, approximately spherical, and a diameter of approximately 30 mm was selected for the following experiments. According to our previous studies, a mixture of 0.1 g SAP and 0.5 ml anhydrous ethanol, and the solution of 0.1 ml UCA SonoVue, or both materials were injected into different cyst before HIFU ablation, respectively. The cyst inoculated with the SAP and UCA alone or in combination was immediately implanted into the abdominal cavity of rabbit for HIFU ablation at a dosage of 100 W acoustic powers. The ablation mode was spot scanning at the speed of 3 mm/s. Every target point was scanned three times; every ablating time lasted 3 s. The distance of each ablated layer was 5 mm. The total ablation time depended on the volume of cyst. The comparison of ultrasound image for each layer of hydatid cyst was made before and after HIFU ablation. The protoscolices in ablated cysts were stained by trypan blue exclusion assay, and their structures were observed by light microscopy. To estimate ablation effects of HIFU to the walls of hydatid cysts, the ultrastructure changes of cyst walls were examined by electron microscopy. The pathological changes of rabbits' skins through which ultrasound penetrated were observed to investigate the side effects of HIFU ablation. The results demonstrated that HIFU had some lethal effects to hydatid cysts in vivo, namely, echo enhancements of ultrasound images of cysts, increases in mortality rate of protoscolices from 15.19 % (HIFU alone) to 48.66 % (HIFU + SAP), 38.67 % (HIFU + UCA), and 67.75 % (HIFU + SAP + UCA), respectively, serious structural damages of protoscolices, and destructions or even disappearance of laminated layers and germinal layers in the walls of hydatid cysts ablated by HIFU aided with UCA and SAP alone or in combination. This study demonstrated that destructive effects of HIFU to transplanted hydatid cyst could be enhanced by UCA and SAP alone, but the destruction of HIFU aided with a combination of UCA and SAP to hydatid cysts was more effective than those aided with UCA or SAP alone. The enhanced thermal and cavitation effects of HIFU induced by UCA and SAP might be involved in the enhanced destructive effects of HIFU on hydatid cysts. There were no evidences of pathological changes on rabbits' skins overlying the hydatid cysts after HIFU ablation. The results suggested that the rabbit model with transplanted hydatid cyst may serve as an optional animal model for the experiments of HIFU ablation to hydatid cyst in vivo, and the materials of UCA and SAP were proved as enhancing agents of HIFU ablation to hydatid cysts, and HIFU at a dosage of 100 W acoustic powers was a safe and feasible parameter to ablate the hydatid cysts in this special animal model. These results laid a theoretical foundation for improving HIFU therapy for cystic echinococcosis by inoculation of UCA and SAP into hydatid cysts.

Development of a biomimetic DNA delivery system by encapsulating polyethyleneimine functionalized silicon quantum dots with cell membranes.

Quantum dots (QDs) are renowned for their remarkable optoelectronic properties, making them suitable for applications such as bioimaging and optoelectronics. However, their use in gene delivery has been restricted due to the low DNA loading capacity. This study aimed to develop a biomimetic DNA delivery system by encapsulating polyethyleneimine (PEI) functionalized silicon QDs (SiQDs) with cell membranes and evaluate its potential as a gene vector in vitro. To achieve this, hydrophilic dispersed silicon QDs (PQDs) were prepared through a one-pot hydrothermal reaction of PEI and 3-Aminopropyltrimethoxysilane (APTMS). Subsequently, red blood cell membrane (RBCM) encapsulated biomimetic QDs (CM-PQDs) was obtained through the extrusion method. The CM-PQDs exhibited higher DNA loading capacity and better stability than naked SiQDs. The CM-PQDs/DNA complex was effectively taken up by cells, as observed through the fluorescence characteristics of QDs themselves. Both CM-P10QDs (prepared with PEI10k) and CM-P25QDs (prepared with PEI25k) could deliver DNA into cells and express the reporter protein successfully. CM-P25QDs showed a higher transfection efficiency of 77.32% in 293 T cells and 47.11% in HeLa cells than SiQDs and CM-P10QDs. The results also indicated that cell membrane encapsulation could effectively reduce the cytotoxicity of SiQDs further. Therefore, the study concludes that CM-PQDs have the potential to serve as a safe and traceable biomimetic gene delivery system.

Super-tough and self-healable all-cellulose-based electrolyte for fast degradable quasi-solid-state supercapacitor.

Recyclable and degradable supercapacitors have promising applications for a sustainable energy storage industry. Herein, we prepare a dual-physical crosslinking (DP) carboxymethyl cellulose (CMC) hydrogel with high-toughness, healability, and electric conductivity by integrating abundant ions into the matrix. The prepared hydrogel displays a maximum compressive fracture stress of 4.42 MPa, fast healing in five seconds, and full degradation within eight days. Moreover, the fabricated supercapacitor shows high specific capacitance (309 F g-1) and volumetric capacitance (2.60 F cm-3). The supercapacitor achieves a healing efficiency of 93.9 % after five cuttings, and exhibits a cycling stability of 84.6 % capacitance retention after 1000 cycles. These merits ensure that the all-cellulose-based supercapacitor can operate in case of sudden collision and deformation, which contribute to reducing the environmental hazards from supercapacitor's preparation to its abandonment.

Early Warning of Infectious Diseases in Hospitals Based on Multi-Self-Regression Deep Neural Network.

Objective: Infectious diseases usually spread rapidly. This study aims to develop a model that can provide fine-grained early warnings of infectious diseases using real hospital data combined with disease transmission characteristics, weather, and other multi-source data. Methods: Based on daily data reported for infectious diseases collected from several large general hospitals in China between 2012 and 2020, seven common infectious diseases in medical institutions were screened and a multi self-regression deep (MSRD) neural network was constructed. Using a recurrent neural network as the basic structure, the model can effectively model the epidemiological trend of infectious diseases by considering the current influencing conditions while taking into account the historical development characteristics in time-series data. The fitting and prediction accuracy of the model were evaluated using mean absolute error (MAE) and root mean squared error. Results: The proposed approach is significantly better than the existing infectious disease dynamics model, susceptible-exposed-infected-removed (SEIR), as it addresses the concerns of difficult-to-obtain quantitative data such as latent population, overfitting of long time series, and considering only a single series of the number of sick people without considering the epidemiological characteristics of infectious diseases. We also compare certain machine learning methods in this study. Experimental results demonstrate that the proposed approach achieves an MAE of 0.6928 and 1.3782 for hand, foot, and mouth disease and influenza, respectively. Conclusion: The MRSD-based infectious disease prediction model proposed in this paper can provide daily and instantaneous updates and accurate predictions for epidemic trends.