Abstract: A tunnel automatic monitoring and measuring equipment and method based on fixed-point tour measurement are provided. The equipment includes a monitoring trolley that can move freely in the longitudinal direction of the tunnel, multiple automatic tracking and identification devices are set on the monitoring trolley, the automatic tracking and identification device is connected to the background processing system telecommunication; a monitoring points with reflective markings are arranged on the surface of the tunnel support structure, under the cooperation of the monitoring trolley and the automatic tracking and identification device, the reflective markings set on the tunnel support structure can be automatically measured at the fixed point to obtain the coordinate information of the relevant monitoring points; then, the deformation data of the support structure required in the construction process are extracted by coordinate information calculation.

Abstract: A real-time fluorescence quantitative RT-qPCR method for direct detection of circulating miRNAs in serum or plasma without the need of extracting nucleic acids. Said method comprises: S1, subjecting cleavages of exosomes and miRNA-protein complexes in serum or plasma, and performing centrifugation to obtain a crude circulating miRNA extract; S2, performing miRNA tailing and reverse transcription; and S3, performing RT-qPCR quantitative detection. Said method does not required nucleic acids to be extracted, and Poly(A) tailing and reverse transcription of miRNA will be synchronously accomplished in one reaction system. The operation is simple, the time is shortened, and the preparation of cDNAs is completed within 95 minutes.

Abstract: Functional shRNA is produced from an expression vector prepared by selecting a two primer design in which the primers are less than about 50 nucleotides in length, annealing and extending the primers using primer extension, digesting the primer extension product and inserting the digestion product into a suitable vector. When the shRNA vectors are inserted into a cell, shRNA transcribed from the vectors modulates gene activity within the cell.

Abstract: A method and primer for quantitative assay of microRNAs (miRNAs) and application of the same are provided. In this method, miRNAs are subjected to polyadenylation and subsequent reverse transcription with miRNA-specific S-Oligo(dT) RT primers. The miRNA is then assayed in a real-time PCR quantitative assay using a miRNA-specific forward primer, a universal reverse primer and a universal probe. The methods in the present invention have improved sensitivity, specificity, and efficiency in miRNA quantification assay, providing a simple, high-throughput, and cost-effective tool applicable for the early diagnosis and prognosis of many severe diseases such as malignancy.

Abstract: A method for amplifying oligonucleotide in vitro by polymerase-endonuclease chain reaction (PECR) which utilizes a single-stranded DNA probe containing repeat sequences, extends a target oligonucleotide by a thermostable DNA polymerase, cleaves extended products with a thermostable endonuclease, and amplifies target oligonucleotide by thermocycling. In PECR, a specific oligonucleotide is exponentially amplified using one single probe instead of a pair of primers, and the reaction is precisely controlled by thermal cycles whose parameters are flexibly adjustable according to length, sequence, melting temperature and initial amount of the target oligonucleotide. Amplification speed depends totally on initial amount of target oligonucleotide present in the reaction system. The method can be used to amplify specific small nucleic acids, such as oligonucleotides and microRNAs, and further conduct quantitative analysis.

Abstract: Functional shRNA is produced from an expression vector prepared by selecting a two primer design in which the primers are less than about 50 nucleotides in length, annealing and extending the primers using primer extension, digesting the primer extension product and inserting the digestion product into a suitable vector. When the shRNA vectors are inserted into a cell, shRNA transcribed from the vectors modulates gene activity within the cell.

Abstract: A research or therapeutic tool for RNA interference (RNAi) is a single vector that expresses multiple short hairpin RNA (shRNA) sequences.

Abstract: Cell-specific methods for silencing genes using double-strand inhibitory RNA (iRNA) are provided, as are constructs for carrying out the methods.