CRISPR/Cas12-Based Ultra-Sensitive and Specific Point-of-Care Detection of HBV.

Hepatitis B remains a major global public health challenge, with particularly high prevalence in medically disadvantaged western Pacific and African regions. Although clinically available technologies for the qPCR detection of HBV are well established, research on point-of-care testing has not progressed substantially. The development of a rapid, accurate point-of-care test is essential for the prevention and control of hepatitis B in medically disadvantaged rural areas. The development of the CRISPR/Cas system in nucleic acid detection has allowed for pathogen point-of-care detection. Here, we developed a rapid and accurate point-of-care assay for HBV based on LAMP-Cas12a. It innovatively solves the problem of point-of-care testing in 10 min, particularly the problem of sample nucleic acid extraction. Based on LAMP-Cas12a, visualization of the assay results is presented by both a fluorescent readout and by lateral flow test strips. The lateral flow test strip technology can achieve results visible to the naked eye, while fluorescence readout can achieve real-time high-sensitivity detection. The fluorescent readout-based Cas12a assay can achieve HBV detection with a limit of detection of 1 copy/µL within 13 min, while the lateral flow test strip technique only takes 20 min. In the evaluation of 73 clinical samples, the sensitivity and specificity of both the fluorescence readout and lateral flow test strip method were 100%, and the results of the assay were fully comparable to qPCR. The LAMP-Cas12a-based HBV assay relies on minimal equipment to provide rapid, accurate test results and low costs, providing significant practical value for point-of-care HBV detection.

Rapid Preparation of Mesoporous Methylsilsesquioxane Aerogels by Microwave Heating Technology.

Microwave heating technology is known as an alternative to traditional gas and electric heating sources. In this work, mesoporous methylsilsesquioxane (MSQ) aerogels were prepared via a sol-gel process accompanied by microwave heating technology, and microwave heating was used in the gelation of sol and the drying of wet gels, respectively. The effects of hexadecyltrimethylammonium chloride (CTAC) as a surfactant and template, hydrochloric acid (HCl) as a catalyst, ethanol as a solvent, sodium hydroxide (NaOH) as a gelation agent, and microwave power on the pore structure of as-prepared MSQ aerogels were investigated in detail. Microwave heating at low power results in the acceleration of sol-gel transition and achieves the gelation within a few minutes. Appropriate amounts of chemical reagents and microwave heating at high power allow the preparation of mesoporous MSQ aerogels with a BET-specific surface area of 681.6 m2·g-1 and a mesopore size of 19 nm, and the resultant MSQ aerogel still has a BET specific surface area as high as 134 m2·g-1 after heat treatment at 600 °C for 2 h, showing high thermal stability. The MSQ aerogels/fibre composite possesses a low thermal conductivity of 0.039 W/(m·k)-1, displaying good thermal insulation. Microwave heating technology is a promising heating method for the preparation of other aerogels.

Extended ORF8 Gene Region Is Valuable in the Epidemiological Investigation of Severe Acute Respiratory Syndrome-Similar Coronavirus.

Severe acute respiratory syndrome coronavirus (SARS-CoV) was discovered as a novel pathogen in the 2002-2003 SARS epidemic. The emergence and disappearance of this pathogen have brought questions regarding its source and evolution. Within the genome sequences of 281 SARS-CoVs, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and SARS-related CoVs (SARSr-CoVs), a ~430 bp genomic region (from 27 701 bp to 28 131 bp in AY390556.1) with regular variations was investigated. This ~430 bp region overlaps with the ORF8 gene and is prone to deletions and nucleotide substitutions. Its complexity suggested the need for a new genotyping method for coronaviruses related to SARS-similar coronaviruses (SARS-CoV, SARSr-CoV, and SARS-CoV-2). Bat SARSr-CoV presented 3 genotypes, of which type 0 is only seen in bat SARSr-CoV, type I is present in SARS in the early phase, and type II is found in all SARS-CoV-2. This genotyping also shows potential usage in distinguishing the SARS-similar coronaviruses from different hosts and geographic areas. This genomic region has important implications for predicting the epidemic trend and studying the evolution of coronavirus.

Fabrication and Characterization of MSQ Aerogel Coating on ePTFE Thin Films for Cable Sheaths.

With methylsilsesquioxane (MSQ) aerogels synthesized by the sol-gel method as a raw material and Si-Ti sol as a binder, an alcohol-based aerogel slurry consisting of only MSQ aerogel and Si-Ti sol was prepared and coated on expanded polytetrafluoroethylene (ePTFE) to form an MSQ aerogel coating layer, followed by low-temperature heat treatment. The effect of Si-Ti sol content on the microstructure of the MSQ aerogel coating layer was investigated, and the properties of a typical MSQ aerogel-layer-coated ePTFE film were evaluated. The results show that Si-Ti sol has an important role in terms of film-forming capability, surface smoothness, flexibility, and powder dropping of the MSQ aerogel coating layer. With a Si-Ti sol of 10.5 wt.% content as a binder and after heat treatment at 170 °C for 30 min, the coated ePTFE flexible thin film with a layer thickness of 30 µm shows high uniformity, integrity, and electrical insulation properties, with an elongation at break decrease over 130%, a thermal conductivity of 0.1753 W/(m·K) at 25 °C, a dielectric constant of 16.5674, and a dielectric loss of 0.06369, which can be promisingly applied in cable sheaths.

Facile Synthesis of Flexible Methylsilsesquioxane Aerogels with Surface Modifications for Sound- Absorbance, Fast Dye Adsorption and Oil/Water Separation.

New flexible methylsilsesquioxane (MSQ) aerogels have been facilely prepared by a sol-gel process with methyltrimethoxysilane (MTMS) and dimethyldimethoxysilane (DMDMS) as co-precursors, followed by surface modification and ambient pressure drying. The microstructure, mechanical properties and hydrophobicity of these MSQ aerogels after surface modifications of hexamethyldisiloxane (HMDSO) and/or hexamethyldisilazane (HMDS) were investigated in detail, and the applications of surface-modified MSQ aerogels in sound-absorbance, fast dye adsorption and oil/water separation were evaluated, respectively. The MSQ aerogels surface-modified by HMDS possess flexibility, elasticity and superhydrophobicity, and demonstrate good performance in the mentioned applications. The resultant MSQ aerogel used in sound-absorbance has high frequency (about 6 kHz) acoustic absorptivity of up to 80%, benefiting from its macroporous structure and porosity of 94%, and it also possesses intermediate frequency acoustic absorptivity (about 1 kHz) up to 80% owing to its elasticity. This MSQ aerogel can selectively separate oil from oil/water mixtures with high efficiency due to its superhydrophobicity and superlipophilicity, resulting from a lot of methyl groups, density as low as 0.12 cm³·g-1 and a water contact angle as high as 157°. This MSQ aerogel can be assembled to be a monolithic column applied for fast dye adsorption, and shows selective adsorption for anionic dyes and removal efficiency of methyl orange of up to 95%.

Facile Preparation of Cellulose Aerogels with Controllable Pore Structure.

Cellulose aerogels are the latest generation of aerogels and have also received extensive attention due to their renewable and biocompatible properties. Herein, cellulose aerogel was facilely prepared by using NaOH/urea solution as solvent, raising the temperature to control gelation and drying wet gel sequentially. With NaOH/urea solution as solvent, the cellulose concentration has an important impact on the micromorphology of cellulose aerogels, while the aging time rarely affects the micromorphology. The appropriate solvent and drying method allow the formation of different cellulose crystalline structures. Different from the Cellulose Ⅰ crystalline structure of raw cellulose powder, the cellulose phase of as-prepared cellulose aerogels belongs to the Cellulose Ⅱ crystalline structure, and to some extent the pyrolysis temperature is also lower than that of raw cellulose powder. The resultant cellulose aerogel prepared by using NaOH/urea solution as solvent and freeze-drying has a uniform macroporous structure with a macropore size of 1~3 µm.

Advances in Field Detection Based on CRISPR/Cas System.

Rapid and accurate diagnostic methods are essential to interrupt outbreaks of infectious diseases such as COVID-19. However, the most commonly used nucleic acid detection method, qPCR or RT-qPCR, takes several hours to complete and requires highly sophisticated equipment. Recently, an emerging nucleic acid detection method based on the CRISPR/Cas system has reduced the reliance on qPCR. It has several important features that make it suitable for on-site POCT (point-of-care testing), including short detection cycles, low cost, high sensitivity, and the ability to be combined with different readout methods. This review briefly introduces the steps of CRISPR/Cas detection and then summarizes the current advances of CRISPR/Cas-based POCT from four steps: nucleic acid extraction, target amplification, CRISPR/Cas-based signal generation, and signal output. Finally, we discuss the advantages and challenges of CRISPR-based POCT and describe the future research perspectives for CRISPR.

Association Between Human Papilloma Virus Infection and Malignant Sinonasal Inverted Papilloma.

OBJECTIVE: This systematic review and meta-analysis aimed to evaluate the risk of malignant sinonasal inverted papilloma (SNIP) according to the type of human papilloma virus (HPV) infection. METHODS: The databases of PubMed, EmBase, and Web of Science were searched for studies that reported the risk of malignant SNIP in patients infected by specific types of HPV. The quantitative analyses for pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. RESULTS: Twenty-six molecular epidemiological studies that recruited a total of 900 patients with SNIP were selected for the final meta-analysis. The summary ORs indicated that HPV-6 (OR: 2.02; 95% CI: 0.47-8.61; P = .343), HPV-11 (OR: 0.86; 95% CI: 0.26-2.89; P = .806), and HPV-6/11 (OR: 1.44; 95% CI: 0.59-3.53; P = .426) infections were not associated with the risk of malignant SNIP. However, the risk of malignant SNIP was increased in patients infected with HPV-16 (OR: 8.51; 95% CI: 3.36-21.59; P < .001), HPV-11/16 (OR: 7.95; 95% CI: 1.13-56.01; P = .038), HPV-18 (OR: 23.26; 95% CI: 5.27-102.73; P < .001), and HPV-16/18 (OR: 24.34; 95% CI: 5.74-103.18; P < .001). CONCLUSIONS: This study found that patients infected with HPV types 16, 11/16, 18, and 16/18 were associated with an increased risk of malignant SNIP. However, patients infected with HPV types 6, 11, and 6/11 did not have a significant risk of malignant SNIP. Laryngoscope, 131:1200-1205, 2021.

Pathological Features of Enterovirus 71-Associated Brain and Lung Damage in Mice Based on Quantitative Proteomic Analysis.

The outbreaks of enterovirus 71 (EV71)-associated hand, foot, and mouth disease (HFMD) have emerged as an emergency of global health due to its association with fatal encephalitis and subsequent neurogenic pulmonary edema; however, the molecular characteristics and pathological features underlying EV71-associated encephalitis and pulmonary edema remain largely unknown. In this study, we performed a proteomic analysis of fresh brain and lung tissues from EV71-infected mice at 7 days post infection. We detected a perturbed expression of 148 proteins in the brain and 78 proteins in the lung after EV71 expression. Further analysis showed that the dysregulated proteins in the brain are involved in a variety of fundamental biological pathways, including complement and coagulation cascades, innate and adaptive immune responses, platelet activation, and nitrogen metabolism, and those proteins in the lung participate in innate and adaptive immune responses, phagosome, arginine biosynthesis, and hypoxia-inducible factor 1 signaling pathway. Our results suggested that immune activation, complement and coagulation dysfunction, platelet activation, imbalance of nitrogen metabolism, and hypoxia could be involved in the pathogenesis of EV71, which explains the major clinical manifestation of hyperinflammatory status of severe HFMD cases. Our study provides further understanding of the molecular basis of EV71 pathogenesis.

CRISPR/Cas System: A Potential Technology for the Prevention and Control of COVID-19 and Emerging Infectious Diseases.

The continued global pandemic of coronavirus disease 2019 (COVID-19) poses a serious threat to global public health and social stability and it has become a serious global public health problem. Unfortunately, existing diagnostic and therapeutic approaches for the prevention and control of COVID-19 have many shortcomings. In recent years, the emerging CRISPR/Cas technology can complement the problems of traditional methods. Biological tools based on CRISPR/Cas systems have been widely used in biomedicine. In particular, they are advantageous in pathogen detection, clinical antiviral therapy, drug, and vaccine development. Therefore, CRISPR/Cas technology may have great potential for application in the prevention and control of COVID-19 and emerging infectious diseases in the future. This article summarizes the existing applications of CRISPR/Cas technology in infectious diseases with the aim of providing effective strategies for the prevention and control of COVID-19 and other emerging infectious diseases in the future.

Facile Synthesis of Methylsilsesquioxane Aerogels with Uniform Mesopores by Microwave Drying.

Methylsilsesquioxane (MSQ) aerogels with uniform mesopores were facilely prepared via a sol⁻gel process followed by microwave drying with methyltrimethoxysilane (MTMS) as a precursor, hydrochloric acid (HCl) as a catalyst, water and methanol as solvents, hexadecyltrimethylammonium chloride (CTAC) as a surfactant and template, and propylene oxide (PO) as a gelation agent. The microstructure, chemical composition, and pore structures of the resultant MSQ aerogels were investigated in detail to achieve controllable preparation of MSQ aerogels, and the thermal stability of MSQ aerogels was also analyzed. The gelation agent, catalyst, solvent, and microwave power have important roles related to the pore structures of MSQ aerogels. Meanwhile, the microwave drying method was found to not only have a remarkable effect on improving production efficiency, but also to be conducive to avoiding the collapse of pore structure (especially micropores) during drying. The resulting MSQ aerogel microwave-dried at 500 W possessed a specific surface area up to 821 m²/g and a mesopore size of 20 nm, and displayed good thermal stability.

Promyelocytic leukemia zinc finger protein activates GATA4 transcription and mediates cardiac hypertrophic signaling from angiotensin II receptor 2.

BACKGROUND: Pressure overload and prolonged angiotensin II (Ang II) infusion elicit cardiac hypertrophy in Ang II receptor 1 (AT(1)) null mouse, whereas Ang II receptor 2 (AT(2)) gene deletion abolishes the hypertrophic response. The roles and signals of the cardiac AT(2) receptor still remain unsettled. Promyelocytic leukemia zinc finger protein (PLZF) was shown to bind to the AT(2) receptor and transmit the hypertrophic signal. Using PLZF knockout mice we directed our studies on the function of PLZF concerning the cardiac specific transcription factor GATA4, and GATA4 targets. METHODOLOGY AND PRINCIPAL FINDINGS: PLZF knockout and age-matched wild-type (WT) mice were treated with Ang II, infused at a rate of 4.2 ng·kg(-1)·min(-1) for 3 weeks. Ang II elevated systolic blood pressure to comparable levels in PLZF knockout and WT mice (140 mmHg). WT mice developed prominent cardiac hypertrophy and fibrosis after Ang II infusion. In contrast, there was no obvious cardiac hypertrophy or fibrosis in PLZF knockout mice. An AT(2) receptor blocker given to Ang II-infused wild type mice prevented hypertrophy, verifying the role of AT(2) receptor for cardiac hypertrophy. Chromatin immunoprecipitation and electrophoretic mobility shift assay showed that PLZF bound to the GATA4 gene regulatory region. A Luciferase assay verified that PLZF up-regulated GATA4 gene expression and the absence of PLZF expression in vivo produced a corresponding repression of GATA4 protein. CONCLUSIONS: PLZF is an important AT(2) receptor binding protein in mediating Ang II induced cardiac hypertrophy through an AT(2) receptor-dependent signal pathway. The angiotensin II-AT(2)-PLZF-GATA4 signal may further augment Ang II induced pathological effects on cardiomyocytes.

Antagonism or synergism. Role of tyrosine phosphatases SHP-1 and SHP-2 in growth factor signaling.

SHP-1 and SHP-2 are two Src homology 2 domain-containing tyrosine phosphatases with major pathological implications in cell growth regulating signaling. They share significant overall sequence identity, but their biological functions are often opposite. SHP-1 is generally considered as a negative signal transducer and SHP-2 as a positive one. However, the precise role of each enzyme in shared signaling pathways is not well defined. In this study, we investigated the interaction of these two enzymes in a single cell system by knocking down their expressions with small interfering RNAs and analyzing the effects on epidermal growth factor signaling. Interestingly, knockdown of either SHP-1 or SHP-2 caused significant reduction in the activation of ERK1/2 but not Akt. Furthermore, SHP-1, SHP-2, and Gab1 formed a signaling complex, and SHP-1 and SHP-2 interact with each other. The interaction of SHP-1 with Gab1 is mediated by SHP-2 because it was abrogated by knockdown of SHP-2, and SHP-2, but not SHP-1, binds directly to tyrosine-phosphorylated Gab1. Together, the data revealed that both SHP-1 and SHP-2 have a positive role in epidermal growth factor-induced ERK1/2 activation and that they act cooperatively rather than antagonistically. The interaction of SHP-1 and SHP-2 may be responsible for previously unexpected novel regulatory mechanism of cell signaling by tyrosine phosphatases.