Advances in the use of nanomaterials for nucleic acid detection in point-of-care testing devices: A review.

The outbreak of the coronavirus (COVID-19) has heightened awareness of the importance of quick and easy testing. The convenience, speed, and timely results from point-of-care testing (POCT) in all vitro diagnostic devices has drawn the strong interest of researchers. However, there are still many challenges in the development of POCT devices, such as the pretreatment of samples, detection sensitivity, specificity, and so on. It is anticipated that the unique properties of nanomaterials, e.g., their magnetic, optical, thermal, and electrically conductive features, will address the deficiencies that currently exist in POCT devices. In this review, we mainly analyze the work processes of POCT devices, especially in nucleic acid detection, and summarize how novel nanomaterials used in various aspects of POCT products can improve performance, with the ultimate aims of offering new ideas for the application of nanomaterials and the overall development of POCT devices.

Effects of growth hormone/estrogen/androgen on COVID-19-type proinflammatory responses in normal human lung epithelial BEAS-2B cells.

BACKGROUND: COVID-19 is a disease caused by SARS-CoV-2, which can cause mild to serious infections in humans. We aimed to explore the effect of growth hormone (GH)/estrogen/androgen in normal human lung epithelial BEAS-2B cells on COVID-19-type proinflammatory responses. METHODS: A BEAS-2B COVID-19-like proinflammatory cell model was constructed. After that, the cells were treated with GH, 17ß-estradiol (E2), and testosterone (Tes) for 24 h. CCK-8 assays were utilized to evaluate cell viability. The mRNA expression of ACE2, AGTR1, TMRRSS2, and ISG15 and the protein expression of ACE2, AGTR1, TMRRSS2, and ISG15 were measured by qRT‒PCR and Western blotting, respectively. ELISAs were performed to determine IL-6, MCP-1, MDA and SOD expression. Flow cytometry was used to measure ROS levels. Finally, MAPK/NF-κB pathway-related factor expression was evaluated. RESULTS: The COVID-19-type proinflammatory model was successfully constructed, and 1000 ng/mL RBD treatment for 24 h was selected as the condition for the model group for subsequent experiments. After RBD treatment, cell viability decreased, the mRNA expression of ACE2, AGTR1, TMRRSS2, and ISG15 and the protein expression of ACE2, AGTR1, TMRRSS2, and ISG15 increased, IL-6, MCP-1, MDA and ROS levels increased, and MDA levels decreased. The mRNA levels of MAPK14 and RELA increased, but the protein levels did not change significantly. In addition, phospho-MAPK14 and phospho-RELA protein levels were also increased. Among the tested molecules, E2 had the most pronounced effect, followed by GH, while Tes showed the opposite effect. CONCLUSION: GH/E2 alleviated inflammation in a COVID-19-type proinflammatory model, but Tes showed the opposite effect.

Characterization of ferroptosis signature to evaluate the predict prognosis and immunotherapy in glioblastoma.

BACKGROUND: Glioblastoma (GBM) is the most common type of brain cancer with poor survival outcomes and unsatisfactory response to current therapeutic strategies. Recent studies have demonstrated that ferroptosis-related genes (FRGs) are linked with the occurrence and development of GBM and may become promising biological indicators in GBM therapy. METHODS: We systematically assessed the relationship between FRGs expression profiles and prognosis in glioma patients based on the Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) datasets to establish a risk score model according to the gene signature of multiple survival-associated DEGs. Further, the differences between the tumor microenvironment score, immune cell infiltration, immune checkpoint expression levels, and drug sensitivity in the high- and low-risk group are analyzed through a variety of algorithms in R software. RESULTS: GBM patients were divided into two subgroups (high- and low-risk) according to the established risk score model. Patients in the high-risk group showed significantly reduced overall survival compared with those in the low-risk group. Also, we found that the high-risk group showed higher ImmuneScore and StromalScore, while different subgroups have significant differences in immune cell infiltration, immune checkpoint expression levels, and drug sensitivity. In summary, we developed and validated an FRGs risk model, which served as an independent prognostic indicator for GBM. Besides, the two subgroups divided by the model have significant differences, which provides novel insights for further studies as well as the personalized treatment of patients.

LncRNA GAS5 regulates epithelial-mesenchymal transition and viability of glioma cells by targeting microRNA-106b and regulating PTEN expression.

LncRNA growth arrest special 5 (GAS5) and microRNA-106b (miR-106b) have been reported to be involved in the regulation of gliomas. However, their precise mechanisms in regulating the progression and development of gliomas remain unclear. We aimed to investigate the interaction between GAS5 and miR-106b, and their influence on the proliferation, migration, and invasion of gliomas cells. Western blotting and qRT-PCR were applied for measuring expression of protein and mRNA, respectively. The proliferation, migration, and invasion of cells were measured by MTT, wound healing, and transwell assays, respectively. Dual luciferase reporter assay was applied for confirming the binding site between miR-106b and GAS5, miR-106b and PTEN. Significant higher expression of miR-106b, and lower expression of GAS5 and PTEN in the glioma tissues were observed. The binding sites between GAS5 and miR-106b, miR-106b and PTEN were identified. GAS5 could regulate the expression of PTEN through targeting miR-106b, and further influence EMT process, and the proliferation, migration, and invasion of gliomas cells. Meanwhile, PTEN could remarkably inhibited the proliferation, migration and invasion of glioma cells. The influence of PTEN on glioma cells and EMT was similar to GAS5. GAS5 could regulate the EMT process, and the migration of gliomas cells through miR-106b targeting PTEN. Therefore, our findings may provide a new thought for the study of pathogenesis and treatment of glioma.

The effect of hypoxia on the proliferation capacity of dermal papilla cell by regulating lactate dehydrogenase.

Hypoxia is of great significance for stem cells to maintain the proliferation and differentiation capacity. As a specialized mesenchymal component of the hair follicle (HF), the dermal papilla cell (DPC) not only regulates HF cycle, but also plays a pivotal role in differentiating hair follicle stem cell(HFSC) into HF. However, whether hypoxia could affect DPCs on proliferation or metabolism remains unclear. In our study, DPCs were cultured in normoxia (20%O2 ) or hypoxia (5%O2 ). Cell viability assays were performed, and lactate dehydrogenase (LDH) activity and lactate level in DPCs were detected. After that, LDH was overexpressed or knocked down in DPCs; then, the expression of protein markers (ALP, Ki-67) was assessed by Western blotting, and cell proliferation was also detected after overexpression or knockdown of LDH. Hypoxia did show positive effect on proliferation of DPCs. The LDH activity of DPCs cultured under hypoxic condition was significantly higher than that of cultured under normoxic condition. Overexpression of LDH significantly up-regulates the expression of ALP and Ki-67 compared with knockdown and negative control. Cell proliferation was also promoted in DPCs with elevated LDH. Our findings showed that the proliferation activity of DPCs could be stimulated under hypoxia. Meanwhile, LDH plays an important role in maintaining the activity of DPCs in hypoxic condition.

Novel Detection of Norovirus and Double Clostridium difficile in a Closed Cartridge System.

Clostridium difficile infection (CDI) has become the main cause of diarrhea-related diseases in domestic (China) inpatients. High-sensitivity and high-specificity detection methods for CDI must be applied clinically for CDI supervisory control. In this paper, we introduce a detection method for C. difficile and Norovirus based on real-time PCR. We developed and optimized a primer-probe for Norovirus targets tcdA and tcdB with remarkably increased detection sensitivity. We then used this method in an integrated cassette, and found increased detection efficiency for Norovirus standards in the cassette compared to C. difficile samples. These results provide a basis for further exploration of automatic testing system design.

Carotid artery calcification score and its association with cognitive impairment.

PURPOSE: To retrospectively investigate the possible association between carotid artery calcification score (CS) and cognitive impairment in carotid artery stenosis (CAS) patients. PATIENTS AND METHODS: Carotid artery was measured in 102 patients with cervical carotid arteries using Color Doppler ultrasound, multi-detector row spiral CT angiography and MRI scanning. Correlation analysis between CSs obtained by MD CT and cognitive scores was performed, and the correlation between CSs and vascular stenosis degree and MRI-measured plaque histological (lipid-rich necrotic nucleus [LRNC], intraplaque hemorrhage and fibrous cap surface rupture) and morphological parameters (lumen area [LA], wall area [WA], total area of blood vessels [TVA], plaque burden [PB]) was analyzed. Follow-up review analysis was conducted on 38 postoperative patients. RESULTS: Significant negative correlation was discovered between CS value and cognitive scores in CAS patients (R=-0.359, P<0.001), which did not exist in postoperative patients (P=0.348); CS value also showed significant correlation with WA (R=0.521, P=0.042), TVA (R=0.215, P=0.017) and PB (R=0.237, P=0.003) and had a certain predictive value for the occurrence probability of carotid plaque LRNC (P=0.029, AUC =0.780) in preoperative patients. CONCLUSION: Carotid artery CSs have significant correlation with cognitive scores, which could be used as risk factor for early screening of cognitive impairment in CAS patients. The possible mechanism may be related to the calcification impact on the plaque burden.

Symmetrical Hierarchical Stochastic Searching on the Line in Informative and Deceptive Environments.

A stochastic point location (SPL) problem aims to find a target parameter on a 1-D line by operating a controlled random walk and receiving information from a stochastic environment (SE). If the target parameter changes randomly, we call the parameter dynamic; otherwise static. SE can be 1) informative (p > 0.5 where p represents the probability for an environment providing a correct suggestion) and 2) deceptive (p <; 0.5). Up till now, hierarchical stochastic searching on the line (HSSL) is the most efficient algorithms to catch static or dynamic parameter in an informative environment, but unable to locate the target parameter in a deceptive environment and to recognize an environment's type (informative or deceptive). This paper presents a novel solution, named symmetrical HSSL, by extending an HSSL binary tree-based search structure to a symmetrical form. By means of this innovative way, the proposed learning mechanism is able to converge to a static or dynamic target parameter in the range of not only 0.6181 <; p <; 1, but also 0 <; p <; 0.382. Finally, the experimental results show that our scheme is efficient and feasible to solve the SPL problem in any SE.