Wei Cui

The ability to interrogate complex biological environments without prior target knowledge is a key challenge in chemical sensing. Conventional methods are limited in discovering previously unidentified biomarkers. To overcome this, we developed CEMs-SELEX (cell extracellular mixtures–systematic evolution of ligands by exponential enrichment), a de novo selection platform that screens for functional DNAzymes responsive to disease-specific CEMs. Applied to tongue squamous cell carcinoma (TSCC), we identified C502, a DNAzyme activated by tumor secretions with high affinity ( K _d = 24.49 nanomolar) and fast catalytic turnover ( K _cat = 47.56 per second). Integrated multiomics and affinity pull-down identified annexin A2 (ANXA2) as its target, and molecular docking with mutagenesis elucidated the recognition mechanism. Engineering C502 into a DNA Walker enabled ultrasensitive detection of ANXA2 in clinical serum and saliva. In 80 individuals, the assay clearly differentiated patients with TSCC from healthy donors, highlighting its potential as a noninvasive diagnostic framework. Collectively, this work closes the loop from functional probe discovery to clinical assay, offering a generalizable platform for noninvasive cancer diagnostics.

Aptamer-based biosensors have been widely constructed and applied to detect diverse targets. Glutathione S-transferase (GST), a pivotal phase II metabolic enzyme, plays a critical role in biotransformation in vivo, and aberrant GST expression is associated with various health risks. Herein, aptamers targeting GST were systematically selected from a randomized single-stranded DNA (ssDNA) library of 79 nucleotides (nt) using a biotinylated GST-immobilized streptavidin agarose (SA) bead SELEX technology. Following rigorous screening across eight rounds, four aptamers with strikingly similar secondary structures emerged. Among these, Seq3 exhibited the highest affinity towards GST and was selected for further optimization. A semi-rational post-SELEX truncation strategy was then employed based on base composition analysis, secondary structure analysis and affinity assessment. This strategy enabled the systematic removal of redundant nucleotides in Seq3 without compromising its affinity, ultimately yielding a truncated aptamer, Seq3-3, which retains its specificity with a compact 39nt length. Building upon Seq3-3, a double-stranded fluorescent aptamer probe was ingeniously designed for the in vitro detection of GST. The detection mechanism hinges on the competitive displacement of the complementary chain from the probe, mediated by the target protein, leading to the separation of the antisense oligonucleotide from the double-stranded complex. This process triggers the restoration of the fluorescence signal, enabling sensitive detection, and the probe exhibits excellent response within a linear range of GST activity ranging from 0 to 1500 U/L. The results show that not only an efficient strategy for screening robust and practicable aptamers but also an ultrahighly sensitive detection platform for GST was established.

(1) Background: To explore the effect of exercise on the formation and recovery of alcoholic liver disease (ALD) and whether the IL-6–p47^phox oxidative–stress axis is involved in that process. (2) Methods: Firstly, 23 six-week-old male C57BL/6J mice were randomly divided into the Con group, ALD group, ALD + NOXI group, ALD + Ex group, and ALD + Ex + NOXI group. The Liber–DeCarli alcoholic liquid diet was used for 6 weeks to establish the ALD mice model, and the Con group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet, and exercise intervention was started after the ALD model establishment and lasted for another 6 weeks, with or without administration of the NOX inhibitor apocynin by intraperitoneal injection on every exercise training day. Secondly, 28 mice were randomly divided into the Sed group, Eth group, Eth + Ex group and Eth + Ex + NOXI group. The Sed group was given the TP4030C control diet. The remaining groups were fed with the TP4030B alcoholic diet and exercise intervention was started synchronously combined with or without administration of intraperitoneal apocynin injections on every exercise training day for 5 weeks. After each individual experiment was accomplished, physiological assessment and biochemical analysis of blood and tissue samples were examined. (3) Results: The levels of TG in serum and IL-6 protein content in liver tissue in the ALD group were significantly increased compared to the Con group (p < 0.05); compared with ALD, p47^phox expression in muscle was increased significantly in the ALD + NOXI group (p < 0.05), and TG in serum decreased in the ALD + Ex group (p < 0.05). TG in serum, AST/ALT ratio, and IL-6 content in both liver and muscle decreased (p < 0.05) in the ALD + Ex + NOXI group with MDA in muscle significantly increased (p < 0.01). The AST/ALT ratio, TG in serum, SOD in liver, and p47^phox in both liver and muscle in the ALD + Ex + NOXI group were significantly decreased compared with the ALD + NOXI group (p < 0.01). Compared with the ALD + Ex group, the liver index and HDL-C levels in serum were decreased (p < 0.05) in the ALD + Ex + NOXI group. The degree of hepatocyte steatosis and inflammatory infiltration were ameliorated after exercise intervention. In the Eth group, the relative epididymal fat content, HDL-C level, and AST/ALT ratio were significantly decreased, and TG and gp91^phox in liver were significantly higher than in the Sed group (p < 0.05, p < 0.01). Compared with the Eth group, the AST/ALT ratio, MDA in the liver, and NOX4 and p47^phox protein expression in the liver were significantly increased, and body weight decreased significantly in the Eth + Ex group (p < 0.05, p < 0.01), as did TG in the liver and MDA in muscle. In the th + Ex + NOXI group, gp91^phox expression in the liver and body weight were significantly decreased (p < 0.05, p < 0.01). In the Eth + Ex + NOXI group, the ratio of AST/ALT and MDA in muscle were increased when compared with the Eth + Ex group, and the protein expression of gp91^phox and p47^phox were much lower (p < 0.01). (4) Conclusions: 6 weeks of exercise intervention during the recovery phase of ALD ameliorates hepatocyte damage and dyslipidemia through the IL-6–p47^phox oxidative–stress axis, and applying a NOX inhibitor in combination could optimize this. However, drinking alcohol during exercise exacerbates dyslipidemia and oxidative stress, with hepatocyte IL-6–p47^phox downregulated.

As a global pandemic, COVID-19 shows no sign of letting up. With the control of the epidemic in China, the proportion of patients with severe and critical diseases being cured and discharged from hospital has increased, and the recovery of COVID-19 patients has become an important issue that urgently needs attention and solutions. By summarizing the exercise rehabilitation strategies and progress of SARS in 2003, this paper analyzed the differences in clinical indicators and recovery characteristics of severe pneumonia caused by the two viruses, and provided comprehensive exercise guidance and intervention strategies for COVID-19 patients for rehabilitation and nursing by referring to the problems and treatment strategies in the rehabilitation and nursing work of SARS. In the post-epidemic period, China will build a multi-dimensional epidemic prevention system by improving the effectiveness of mass training and strengthening local risk prevention and control. This paper discusses the exercise rehabilitation strategy of SARS patients after recovery, which has guiding significance for exercise intervention and scientific fitness of COVID-19 patients after recovery during epidemic prevention period.