Herein, we have designed a unique oligonucleotide probe named as bumped switchable molecular probe (BS-MB) and employ it to ultrasensitive detection of miRNA-21. The uniqueness is that the bumped portion renders the BS-MB great design flexibility and divides its long stem into two short stems so that it can play the roles of recognition unit, reporting unit, and polymerization primer and template simultaneously. The opening of BS-MB requires a delayed reaction, allowing the system a better-suppressed background without miRNA-21. In contrast, the introduction of miRNA-21 delayed the full opening of BS-MB, inducing a concurrent three-stage amplification for consuming lots of BS-MBs in one amplification event. During the amplification, lots of target analogues of extended miRNA-21 (E-miRNA-21) and cleavaged miRNA-21 (C-miRNA-21) that can be reused as miRNA-21 to cause further fluorescence enhancement are repeatedly generated. The system retains the simplicity (one oligonucleotide probe) and simplifies the detection procedure (one-step detection). Its mix-to-signaling and oneto-many amplification behaviors are superior to conventional MB based amplification, exhibiting a wide linear response and a low detection limit of 8.1 fM. High specificity against even one-base mutation, desirable recovery rates for testing miRNA-spiked serum samples, and well-defined accuracy for classifying clinically related samples are also demonstrated.

Hematological and serum biochemical reference values for 33 free-ranging sambar deer following immobilization with medetomidine (70-100 mu gkg(-1)) and ketamine (1-2 mgkg(-1)) in Sariska Tiger Reserve in India were established and the differences across sex and age group were compared. Hematological and biochemical variables across different age-classes for red blood cell count, eosinophils, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, triglycerides, and blood glucose varied significantly though the hematological values did not vary between sexes. Blood urea nitrogen and creatinine levels were significantly influenced by sex in the present study. Interpretation of the result, however, needs to be made with caution due to the low number of sub-adult compared to adults in the present study. The study highlights the need for understanding the source of individual variation for meaningful interpretation. This paper provides the first report of baseline hematologic and serum biochemistry values for free-ranging sambar deer that can form basis for assessing the state of health and nutritional condition of the animal.