Structural Characteristics of Polysaccharide GP2a in Gardenia jasminoides and Its Immunomodulatory Effect on Macrophages.

Here, we elucidated the structural characteristics of a polysaccharide isolated from Gardenia jasminoides Ellis (labeled as GP2a) and its immunomodulatory activity. GP2a is an acidic polysaccharide with a molecular weight of 44.8 kDa, mostly comprising galacturonic acid. Methylation analysis revealed 4-GalpA (74.8%) to be the major sugar residue in GP2a. Nuclear magnetic resonance analysis indicated that its main chain comprised →4)-α-D-GalpA-6-OMe-(1→4)-α-D-GalpA-(1→ and →4)-α-D-GalpA-6-OMe-(1→2)-α-L-Rhap-(1→, with galactan and arabinans linked to the C-4 position of →2)-α-L-Rhap-(1→ residue as branched chains. Furthermore, GP2a showed no obvious toxicity to macrophages (RAW 264.7) while enhancing cell viability in a dose- and time-dependent manner. Compared with untreated cells, nitric oxide production and secretion of cytokines, such as tumor necrosis factor-α, interferon-γ, interleukin (IL)-1ß, IL-6, and granulocyte macrophage colony stimulating factor, in GP2a-treated cells significantly increased after 48 h. At 300 µg/mL GP2a concentration, there was no significant difference in the cytokine levels in GP2a- and lipopolysaccharide-treated cells (the positive control). In summary, GP2a is a pectic polysaccharide with homogalacturonan and rhamnogalacturonan-I structural regions in the main chain. Based on its immunomodulatory effects in vitro, GP2a may have potential uses in functional food and medicine.

Molecular cloning, characterization and functional analysis of GluCl from the oriental armyworm, Mythimna separata Walker.

Glutamate-gated chloride channels (GluCls) mediate inhibitory synaptic transmission in invertebrate nervous systems, and only one GluCl gene has been found in insects. Therefore, insect GluCls are one of the major targets of insecticides including avermectins. In the present study, a 1347 bp full-length cDNA encoding a 449-amino acid protein (named MsGluCl, GenBank ID: MK336885) was cloned from the oriental armyworm, Mythimna separata, and characterized two alternative splicing variants of MsGluCl. The protein shares 76.9-98.6% identity with other insect GluCl isoforms. Spatial and temporal expression analysis revealed that MsGluCl was highly expressed in the 3rd instar and adult head. Dietary ingestion of dsMsGluCl significantly reduced the mRNA level of MsGluCl and decreased abamectin mortality. Thus, our results reveal that MsGluCl could be the molecular target of abamectin and provide the basis for further understanding the resistance mechanism to abamectin in arthropods.

Silence of ryanodine receptor gene decreases susceptibility to chlorantraniliprole in the oriental armyworm, Mythimna separata Walker.

The ryanodine receptors of insects are the main target sites of diamide insecticides, which show highly selective insecticidal activity relative to toxicity in mammals and provide a novel option for managing lepidopteran pests. The oriental armyworm, Mythimna separata (Walker), is a destructive pest of agricultural crops, and great efforts have been undertaken to control this pest including repeated insecticide applications. In this study, full-length cDNA of a ryanodine receptor gene from M. separata (MsRyR) was cloned and characterized. The cDNA of MsRyR had a 15,372 bp open reading frame and encoded 5124 amino acids (GenBank ID: MG712298). MsRyR shares 78-97% identity with RyR isoforms of other insects, and <50% identities with Homo sapiens RyRs 1-3. Temporal and spatial expression analysis detected MsRyR at all developmental stages and in all tissues. The highest relative levels of MsRyR were detected in the second instar and head. Exposure to chlorantraniliprole after 24 h significantly increased the expression levels of whole body MsRyR mRNA. In addition, dietary ingestion of dsMsRyR significantly reduced the mRNA level of MsRyR and greatly decreased chlorantraniliprole-induced mortality. Our results revealed that the MsRyR could be the molecular target of chlorantraniliprole, and provided the basis for further understanding the resistance mechanism of chlorantraniliprole.

Identification of miRNAs and their targets through high-throughput sequencing and degradome analysis in male and female Asparagus officinalis.

BACKGROUND: MicroRNAs (miRNAs), a class of non-coding small RNAs (sRNAs), regulate various biological processes. Although miRNAs have been identified and characterized in several plant species, miRNAs in Asparagus officinalis have not been reported. As a dioecious plant with homomorphic sex chromosomes, asparagus is regarded as an important model system for studying mechanisms of plant sex determination. RESULTS: Two independent sRNA libraries from male and female asparagus plants were sequenced with Illumina sequencing, thereby generating 4.13 and 5.88 million final clean reads, respectively. Both libraries predominantly contained 24-nt sRNAs, followed by 21-nt sRNAs. Further analysis identified 154 conserved miRNAs, which belong to 26 families, and 39 novel miRNA candidates seemed to be specific to asparagus. Comparative profiling revealed that 63 miRNAs exhibited significant differential expression between male and female plants, which was confirmed by real-time quantitative PCR analysis. Among them, 37 miRNAs were significantly up-regulated in the female library, whereas the others were preferentially expressed in the male library. Furthermore, 40 target mRNAs representing 44 conserved and seven novel miRNAs were identified in asparagus through high-throughput degradome sequencing. Functional annotation showed that these target mRNAs were involved in a wide range of developmental and metabolic processes. CONCLUSIONS: We identified a large set of conserved and specific miRNAs and compared their expression levels between male and female asparagus plants. Several asparagus miRNAs, which belong to the miR159, miR167, and miR172 families involved in reproductive organ development, were differentially expressed between male and female plants, as well as during flower development. Consistently, several predicted targets of asparagus miRNAs were associated with floral organ development. These findings suggest the potential roles of miRNAs in sex determination and reproductive developmental processes in asparagus.

The association between asymmetric hip mobility and neck pain in young adults.

OBJECTIVE: The objective of this cross-sectional observational study was to determine whether asymmetric hip mobility was associated with neck pain in young adults. METHODS: Three hundred twenty-seven freshmen students were recruited from an urban university and underwent the Patrick's flexion, abduction, external rotation, extension (FABERE) test for comparison of the functional mobility of bilateral hip joints during the health examination. A logistic regression model was constructed to determine whether the asymmetry measured by the Patrick's FABERE test was associated with neck pain after adjusting for factors of sex and exercise habits. RESULTS: The frequency of asymmetric results of the Patrick's FABERE test among the students who reported neck pain was significantly higher than that of those without neck pain (54.2% vs 26.5%; P < .001). After adjusting for the above confounders, the odds ratio of asymmetric results of a Patrick's FABERE test was 2.99 (95% confidence interval, 1.57-5.72; P < .001). CONCLUSIONS: Imbalanced mobility of the hip joints might be associated with an increased incidence of neck pain.