A mutation-sensitive, multiplexed and amplification-free detection of nucleic acids by stretching single-molecule tandem hairpin probes.

The detection of nucleic acid sequences in parallel with the discrimination of single nucleotide variations (SNVs) is critical for research and clinical applications. A few limitations make the detection technically challenging, such as too small variation in probe-hybridization energy caused by SNVs, the non-specific amplification of false nucleic acid fragments and the few options of dyes limited by spectral overlaps. To circumvent these limitations, we developed a single-molecule nucleic acid detection assay without amplification or fluorescence termed THREF (hybridization-induced tandem DNA hairpin refolding failure) based on multiplexed magnetic tweezers. THREF can detect DNA and RNA sequences at femtomolar concentrations within 30 min, monitor multiple probes in parallel, quantify the expression level of miR-122 in patient tissues, discriminate SNVs including the hard-to-detect G-U or T-G wobble mutations and reuse the probes to save the cost. In our demonstrative detections using mock clinic samples, we profiled the let-7 family microRNAs in serum and genotyped SARS-CoV-2 strains in saliva. Overall, the THREF assay can discriminate SNVs with the advantages of high sensitivity, ultra-specificity, multiplexing, reusability, sample hands-free and robustness.

Synthesis and biological evaluation of water-soluble derivatives of chiral gossypol as HIV fusion inhibitors targeting gp41.

A series of novel or known water-soluble derivatives of chiral gossypol were synthesized and screened in vitro for their anti-HIV-1 activity. (-)-gossypol derivative was more active against HIV-1 than the corresponding (+)-gossypol derivative, respectively. Among these derivatives, d-glucosamine derivative of (-)-gossypol, oligopeptide derivative of (-)-gossypol and taurine derivative of (-)-gossypol, such as compounds 1a, 3a and 14a, showed significant inhibitory activities against HIV-1 replication, HIV-1 mediated cell-cell fusion and HIV gp41 6-helix bundle formation as some amino acid derivatives of (-)-gossypol.

Study on a novel omnidirectional ultrasonic cavitation removal system for Microcystis aeruginosa.

Microcystis aeruginosa, as a typical alga, produces microcystin with strong liver toxicity, seriously endangering the liver health of human and animals. Inhibiting the bloom of the Microcystis aeruginosa in lakes becomes a significant and meaningful work. Ultrasonic cavitation is currently considered to be the most environmentally friendly and effective method for the removal of Microcystis aeruginosa. However, the commercialized ultrasonic algae removal systems require multi-Langevin transducers to achieve omnidirectional ultrasonic irradiation due to the single irradiation direction of the Langevin transducer, resulting in the complex design and high energy consumption. To achieve a low-cost, simple structure, and high-efficiency algae removal system, a novel omnidirectional ultrasonic cavitation removal system for Microcystis aeruginosa is proposed. The proposed system is major composed of a novel omnidirectional ultrasonic transducer, which generates the omnidirectional ultrasonic irradiation by its shaking-head motion coupled by two orthogonal bending vibration modes. Modal simulation, sound field simulation, and cavitation bubble radius simulation are first carried out to optimize the geometric sizes of the proposed transducer and verify the correctness of the omnidirectional ultrasonic irradiation principle. Then the vibration characteristics of the transducer prototype are measured by vibration tests and impedance tests. Finally, the feasibility and effectiveness of the proposed omnidirectional ultrasonic removal system for Microcystis aeruginosa are evaluated through the algae removal experiments. The experimental results exhibit that the algal cells damaged by ultrasonic irradiation from the proposed system do not have the ability to self-repair. In addition, the algal removal rates reached 55.41% and 72.97% after 30 min of ultrasonic treatment when the corresponding ultrasonic densities are 0.014 W/mL and 0.021 W/mL, respectively. The proposed omnidirectional ultrasonic algae removal system significantly simplifies the configuration and reduces energy consumption, presenting the potential promise of algae removal and environmental protection.

Genetic analysis and molecular detection of resistance to chlorpyrifos mediated by the A216S substitution in acetylcholinesterase-1 in the plant bug Apolygus lucorum.

The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China. Previously, we reported that chlorpyrifos resistance in a laboratory-selected strain of A. lucorum (BZ-R) is associated with the homozygosis of an allele in the ace-1 gene encoding an alanine to serine substitution at position 216 of acetylcholinesterase-1. Here we describe the results of crosses between the resistant BZ-R strain (41-fold to chlorpyrifos) and the unselected susceptible BZ-S strain homozygous for the wild type alanine allele at position 216. Resistance to chlorpyrifos was inherited as a semi-dominant trait mainly controlled by a single autosomal gene and co-segregates strongly but not completely with the serine substitution in ace-1. Synergism bioassays and enzyme assays showed that minor contributions to resistance are also made by enhanced cytochrome P450 and carboxylesterase activities. A survey of 25 field populations from five Chinese provinces showed strong positive correlations between 50% lethal concentration against chlorpyrifos and S216 allele and genotype frequencies, although the most tolerant populations still only show 40%-50% S216 allele frequencies. The results above provide important information for designing effective resistance monitoring and management strategies for A. lucorum in China.

Identification of the ryanodine receptor mutation I4743M and its contribution to diamide insecticide resistance in Spodoptera exigua (Lepidoptera: Noctuidae).

Insect ryanodine receptors (RyRs) are the targets of diamide insecticides. Two point mutations G4946E and I4790M (numbering according to Plutella xylostella, PxRyR) in the transmembrane domain of the insect RyRs associated with diamide resistance have so far been identified in three lepidopteran pests, P. xylostella, Tuta absoluta and Chilo suppressalis. In this study, we identified one of the known RyR target site resistance mutations (I4790M) in a field-collected population of Spodoptera exigua. The field-collected WF population of S. exigua exhibited 154 fold resistance to chlorantraniliprole when compared with the susceptible WH-S strain. Sequencing the transmembrane domains of S. exigua RyR (SeRyR) revealed that the resistant WF strain was homozygous for the I4743M mutation (corresponding to I4790M in PxRyR), whereas the G4900E allele (corresponding to G4946E of PxRyR) was not detected. The 4743M allele was introgressed into the susceptible WH-S strain by crossing WF with WH-S, followed by three rounds of backcrossing with WH-S. The introgressed strain 4743M was homozygous for the mutant 4743M allele and shared about 94% of its genetic background with that of the recipient WH-S strain. Compared with WH-S, the near-isogenic 4743M strain showed moderate levels of resistance to chlorantraniliprole (21 fold), cyantraniliprole (25 fold) and flubendiamide (22 fold), suggesting that the I4743M mutation confers medium levels of resistance to all three diamides. Genetic analysis showed diamide resistance in the 4743M strain was inherited as an autosomal and recessive trait. Results from this study have direct implications for the design of appropriate resistance monitoring and management practices to sustainably control S. exigua.

Association between single nucleotide polymorphism of IL15RA gene with susceptibility to ossification of the posterior longitudinal ligament of the spine.

BACKGROUND: Previous studies reported the association between single nucleotide polymorphism (SNP) of IL15 receptor alpha (IL15RA) gene with susceptibility to ossification of the posterior longitudinal ligament of the spine (OPLL). However, the results were still in controversy. Therefore, the purpose of the present study was to investigate the association between SNPs of IL15RA gene with susceptibility to OPLL in a Chinese Han population. METHODS: A total of 235 OPLL patients and 250 age-matched healthy controls were recruited. All the subjects were genotyped using the PCR (polymerase chain reaction)-based invader assay. A case-control study was performed to define the contribution of rs2228059 and rs2296139 to predisposition of OPLL. We also performed subgroup analysis according to the different gender. RESULTS: A significant association of rs2228059 with OPLL was observed in the Chinese Han population (p <0.001, OR = 1.63, 95% CI = 1.26-2.11). The subgroup analysis showed that there was a significant association between the allele frequency of rs2228059 and the susceptibility of OPLL in males (p = 0.002, OR = 1.72, 95% CI = 1.23-2.42). However, there was no significant association between SNP of rs2296139 and susceptibility to OPLL. CONCLUSIONS: The present study demonstrates that the SNP of rs2228059 in IL15RA gene is associated with susceptibility to OPLL in a Chinese Han population, especially in males.

Heat shock cognate 71 (HSC71) regulates cellular antiviral response by impairing formation of VISA aggregates.

In response to viral infection, RIG-I-like RNA helicases detect viral RNA and signal through the mitochondrial adapter protein VISA. VISA activation leads to rapid activation of transcription factors IRF3 and NF-κB, which collaborate to induce transcription of type I interferon (IFN) genes and cellular antiviral response. It has been demonstrated that VISA is activated by forming prion-like aggregates. However, how this process is regulated remains unknown. Here we show that overexpression of HSC71 resulted in potent inhibition of virus-triggered transcription of IFNB1 gene and cellular antiviral response. Consistently, knockdown of HSC71 had opposite effects. HSC71 interacted with VISA, and negatively regulated virus-triggered VISA aggregation. These findings suggest that HSC71 functions as a check against VISA-mediated antiviral response.

Measuring factors affecting grip strength in a Taiwan Chinese population and a comparison with consolidated norms.

We inquire whether assessment of an individual's upper limb function may be improved by using specific regional norms rather than consolidated global norms. Grip strengths were measured in a sample of 482 adults across Taiwan, and compared with consolidated norms. To ensure comparable conditions, our procedures were those recommended by the American Society of Hand Therapists (ASHT). Overall the mean grip strength of our sample was significantly (male 25%, female 27%) lower than consolidated norms derived from largely Caucasian populations. We investigated variables that might relate to this divergence. Results of ANOVA and stepwise multiple regression analysis showed that gender, age and palm length were effective predictors in grip strength. A regression equation was derived. When other variables were matched, palm length appeared an important discriminating factor. Further anthropometric and socio-economic factors also need investigation. Specific regional norms should provide more accuracy for ergonomists and health workers assessing an individual's upper limb function, and may avoid errors in appraisal. This paper suggests grip strength values for Taiwan.