Summary. -The occurrence of the primer-independent cDNA synthesis during RT-PCR analysis of human and animal RNA viruses has been well documented. Conversely, there is scant knowledge about this event in plant RNA viruses. Here we show that the primer-independent cDNA synthesis occurs in all eight different plant RNA viruses tested in this study, suggesting a common phenomenon for RT-PCR analysis of plant RNA viruses. Additional experiments indicate that the event is likely contributed to by RNA self-priming, and can be effectively reduced or eliminated through increasing temperature of the RT reaction.Keywords: RT-PCR; primer-independent cDNA synthesis; self-priming; plant RNA virus; RT temperature * Corresponding author. E-mail: liweimin01@caas.cn; phone: +86-10-82106117. C. Zhang. and H.N. Wu contributed equally to this work. Abbreviations: CGMMV = cucumber green mottle mosaic virus; CMV = cucumber mosaic virus; gRNA = genomic RNA; ORSV = odontoglossum ring-spot virus; PVX = potato virus X; TCV = turnip crinkle virus; TMV U1 = tobacco mosaic virus U1 strain; TNV = tobacco necrosis virus; TRV = tobacco rattle virus RT-PCR is one of the most common methods of molecular biology used to detect and quantify RNA expression levels in both cells and small quantities of tissues. Within this process, RT is the first and fundamental step, in which the RNA template is converted into its cDNA by reverse transcriptase in the presence of exogenous oligonucleotide referred to as primer, providing template DNA for subsequent PCR amplification. The primer-dependent mechanism for RT reaction has been generally accepted over the years. However, during RT-PCR analysis of human and animal RNA viruses, bacterial operons, as well as eukaryotic cellular RNAs, it has been observed that cDNA could be synthesized by reverse transcriptase without the addition of exogenous oligonucleotide (Gunji et al

Cardiovascular disease morbidity/mortality are increasing due to an aging population and the rising prevalence of diabetes and obesity. Therefore, innovative cardioprotective measures are required to reduce cardiovascular disease morbidity/mortality. The role of necroptosis in myocardial ischemia–reperfusion injury (MI–RI) is beyond doubt, but the molecular mechanisms of necroptosis remain incompletely elucidated. Growing evidence suggests that MI–RI frequently results from the superposition of multiple pathways, with autophagy, ferroptosis, and CypD-mediated mitochondrial damage, and necroptosis all contributing to MI–RI. Receptor-interacting protein kinases (RIPK1 and RIPK3) as well as mixed lineage kinase domain-like pseudokinase (MLKL) activation is accompanied by the activation of other signaling pathways, such as Ca2+/calmodulin-dependent protein kinase II (CaMKII), NF-κB, and JNK-Bnip3. These pathways participate in the pathological process of MI–RI. Recent studies have shown that inhibitors of necroptosis can reduce myocardial inflammation, infarct size, and restore cardiac function. In this review, we will summarize the molecular mechanisms of necroptosis, the links between necroptosis and other pathways, and current breakthroughs in pharmaceutical therapies for necroptosis.