A multiplex direct PCR method for the rapid and accurate discrimination of three species of spider mites (Acari: Tetranychidae) in fruit orchards in Beijing.

Spider mites (Acari: Tetranychidae) are polyphagous pests of economic importance in agriculture, among which the two-spotted spider mite Tetranychus urticae Koch has spread widely worldwide as an invasive species, posing a serious threat to fruit tree production in China, including Beijing. The hawthorn spider mite, Amphitetranychus viennensis Zacher, is also a worldwide pest of fruit trees and woody ornamental plants. The cassava mite, Tetranychus truncatus Ehara, is mainly found in Asian countries, including China, Korea and Japan, and mainly affects fruit trees and agricultural crops. These three species of spider mites are widespread and serious fruit tree pests in Beijing. Rapid and accurate identification of spider mites is essential for effective pest and plant quarantine in Beijing orchard fields. The identification of spider mite species is difficult due to their limited morphological characteristics. Although the identification of insect and mite species based on PCR and real-time polymerase chain reaction TaqMan is becoming increasingly common, DNA extraction is difficult, expensive and time-consuming due to the minute size of spider mites. Therefore, the objective of this study was to establish a direct multiplex PCR method for the simultaneous identification of three common species of spider mites in orchards, A. viennensis, T. truncatus and T. urticae, to provide technical support for the differentiation of spider mite species and phytosanitary measures in orchards in Beijing. Based on the mitochondrial cytochrome c oxidase subunit I (COI) of the two-spotted spider mite and the cassava mite and the 18S gene sequence of the hawthorn spider mite as the amplification target, three pairs of specific primers were designed, and the primer concentrations were optimized to establish a direct multiplex PCR system for the rapid and accurate discrimination of the three spider mites without the need for DNA extraction and purification. The method showed a high sensitivity of 0.047 ng for T. truncatus and T. urticae DNA and 0.0002 ng for A. viennensis. This method eliminates the DNA extraction and sequencing procedures of spider mite samples, offers a possibility for rapid monitoring of multiple spider mites in an integrated microarray laboratory system, reducing the time and cost of leaf mite identification and quarantine monitoring in the field.

[Analysis on regional homogeneity of resting brain during balance acupuncture-induced analgesic effect in migraine patients without aura].

OBJECTIVE: To observe the relationship between the analgesic effect of balance acupuncture and functional changes in brain in patients with migraine without aura. METHODS: A total of 40 cases of migraine without aura were equally randomized into a headache-acupoint group and a sham-acupoint group. When acupuncture given, a filiform needle was inserted into the headache-acupoint (the midpoint of the depression region anterior to the juncture of the first and second metatarsal bones on the dorsum of the foot) or the sham point (the midpoint of the depression region anterior to the juncture site between the 3rd and 4th metatarsal joints of the dorsum of the foot) about 25-40 mm deep and manipulated for a while till the patient experienced feelings of electric shock and numbness, then withdrawn immediately. The treatment was conducted once daily for 4 weeks. The visual analogue scale (VAS) was used to evaluate the severity of pain, and the regional homogeneity (ReHo) analysis of resting state functional magnetic resonance imaging (fMRI) was used to assess changes of the spontaneous brain activity. RESULTS: After acupuncture, the analgesic effect of headache-acupoint was better than that of the sham-acupoint in both intervention stage and the follow-up stage (P< 0.05), and was also stronger in the intervention stage than in the follow-up stage (P<0.05). There was no significant difference in the analgesic effect between the intervention stage and the follow-up stage in the sham-acupoint group (P>0.05). Compared with pre-intervention, 4-weeks' intervention at the headache-acupoint showed an increase of ReHo values in the anterior cingulate gyrus, anterior central gyrus, superior orbital frontal gyrus, insula, inferior lobule, left anterior cingulate gyrus, ventral lateral nucleus and ventral posteromedial nucleus of the thalamus, pontine nucleus, cerebellar tonsils and orbital frontal inferior gyrus of the brain (P<0.05), and a decrease of ReHo values in the right brain bridge, central posterior gyrus, posterior cingulate gyrus, left central anterior gyrus, posterolateral nucleus of thalamus, and hippocampus (P<0.05), separately. In the sham-acupoint group, the ReHo value was increased in the right tongue gyrus, the left anterior lobe, the anterior cingulate gyrus and the lower occipital gyrus of the brain (P<0.05), and reduced in the left ventral posterolateral nucleus of the thalamus, separately (P<0.05). CONCLUSION: Balance acupuncture stimulation of headache acupoint has an analgesic effect in migraine patients without aura, which may be related to its effect in regulating resting state brain function of the limbic-system-dominated multiple brain regions.

Number of and distance between response elements in Kaposi's sarcoma-associated herpesvirus ORF57 promoter influence its activation by replication and transcription activator and its repression by interferon regulatory factor 7.

Kaposi's sarcoma-associated herpesvirus ORF57 expression is highly responsive to replication and transcription activator (RTA) and interferon regulatory factor 7 (IRF-7). Three RTA response elements (RREs) have been identified in the ORF57 promoter. Here, we show evidence of another functional RRE located between nt 82003 and 82081, which can complement the loss of RTA activation resulting from RRE1 deletion. Repeats of a recombination signal-binding protein Jkappa (RBP-Jkappa) site enhanced RTA activation, which could not be suppressed by IRF-7. Alteration of the distance between the RBP-Jkappa site and RRE2 modulated responsiveness to RTA and IRF-7. These results will help to elucidate the precise regulation of viral gene expression.