RG108

J-Lat cells are derivatives of the Jurkat CD4⁺ T cell line that contain a non-infectious, inducible HIV provirus with a GFP tag. While these cells have substantially advanced our understanding of HIV latency, their use by many laboratories in low and middle-income countries is restricted by limited access to flow cytometry. To overcome this barrier, we describe a modified J-Lat assay using a standard microplate reader that detects HIV-GFP expression following treatment with latency-reversing agents (LRAs). We show that HIV reactivation by control LRAs like prostratin and romidepsin is readily detected with dose dependence and with significant correlation and sensitivity to standard flow cytometry. For example, 10 μM prostratin induced a 20.1 ± 3.3-fold increase in GFP fluorescence in the microplate reader assay, which corresponded to 64.2 ± 5.0% GFP-positive cells detected by flow cytometery. Similarly, 0.3 μM prostratin induced a 1.7 ± 1.2-fold increase compared to 8.7 ± 5.7% GFP-positive cells detected. Using this method, we screen 79 epigenetic modifiers and identify CUDC-101, molibresib, and quisinostat as novel LRAs. This microplate reader-based method offers accessibility to researchers in resource-limited regions to work with J-Lat cells and more actively participate in global HIV cure research efforts.

Type 2 diabetes is on the rise year by year, and 80% of patients with type 2 diabetes die from cardiovascular complications.Strict control of blood glucose can significantly reduce the occurrence and development of diabetic microvascular complications, but can not effectively prevent the occurrence of diabetic macrovascular lesions.DNA methyltransferase 1 (DNMT1) plays an important role in developing phenotypic switching and secretion of inflammatory factors in vascular smooth muscle cells(VSMC), leading to the deregulation of vascular function.In this work, a biosensor based on methylation protection was designed.In the presence of the target DNMT 1, the formed methylation sites protect the dsDNA from being digested by the HpaII restriction enzyme.At the same time, a large number of G-quadruplex are produced by the catalyzed hairpin assembly (CHA) reaction cycle, and the peroxidase-like activity of G-tetraplex and hemin is used to continuously catalyze the color change of substrate ABTS, thus realizing dual signal amplification.The binding of magnetic beads to dsDNA effectively reduces the background signal and enables a highly selective and sensitive anal. of the target DNMT 1 in complex biol. samples, with a detection limit as low as 0.22 UmL^-1, showing a good linear relationship in the range of 0.22-10 UmL^-1.A new approach for the study of macrovascular damage is provided in type 2 diabetes.