A Regulatory Mechanism on Pathways: Modulating Roles of MYC2 and BBX21 in the Flavonoid Network.

Genes of metabolic pathways are individually or collectively regulated, often via unclear mechanisms. The anthocyanin pathway, well known for its regulation by the MYB/bHLH/WDR (MBW) complex but less well understood in its connections to MYC2, BBX21, SPL9, PIF3, and HY5, is investigated here for its direct links to the regulators. We show that MYC2 can activate the structural genes of the anthocyanin pathway but also suppress them (except F3'H) in both Arabidopsis and Oryza when a local MBW complex is present. BBX21 or SPL9 can activate all or part of the structural genes, respectively, but the effects can be largely overwritten by the local MBW complex. HY5 primarily influences expressions of the early genes (CHS, CHI, and F3H). TF-TF relationships can be complex here: PIF3, BBX21, or SPL9 can mildly activate MYC2; MYC2 physically interacts with the bHLH (GL3) of the MBW complex and/or competes with strong actions of BBX21 to lessen a stimulus to the anthocyanin pathway. The dual role of MYC2 in regulating the anthocyanin pathway and a similar role of BBX21 in regulating BAN reveal a network-level mechanism, in which pathways are modulated locally and competing interactions between modulators may tone down strong environmental signals before they reach the network.

Linking New Alleles at the Oscillator Loci to Flowering and Expansion of Asian Rice.

The central oscillator is believed to be the key mechanism by which plants adapt to new environments. However, impacts from hybridization, the natural environment, and human selection have rarely been assessed on the oscillator of a crop. Here, from clearly identified alleles at oscillator loci (OsCCA1/LHY, OsPRR95, OsPRR37, OsPRR59, and OsPRR1) in ten diverse genomes of Oryza sativa, additional accessions, and functional analysis, we show that rice's oscillator was rebuilt primarily by new alleles from recombining parental sequences and subsequent 5' or/and coding mutations. New alleles may exhibit altered transcript levels from that of a parental allele and are transcribed variably among genetic backgrounds and natural environments in RIL lines. Plants carrying more expressed OsCCA1_a and less transcribed OsPRR1_e flower early in the paddy field. 5' mutations are instrumental in varied transcription, as shown by EMSA tests on one deletion at the 5' region of highly transcribed OsPRR1_a. Compared to relatively balanced mutations at oscillator loci of Arabidopsis thaliana, 5' mutations of OsPRR37 (and OsCCA1 to a less degree) were under negative selection while those of OsPRR1 alleles were under strong positive selection. Together, range expansion of Asian rice can be elucidated by human selection on OsPRR1 alleles via local flowering time-yield relationships.

The New Media Environment Presents Challenges and Opportunities for Music Education in Higher Education.

Educational reform is essential. The revolution of multimedia technology and network technology is having a profound impact on the traditional teaching system, teaching methods, and training content. The learner-centered online learning model is rapidly developing. Research shows that, based on new media concepts and technology applications, music teaching platforms can be created and researched through limited forms of music teaching in colleges and universities and new media platforms. The survey results of the practice and role of new media in the process of assisting music teaching in colleges and universities show that most students have a positive attitude towards the new media teaching assistant method in public music courses in colleges and universities and are very interested in this method. Students' aesthetic, analytical, expressive, and collaborative abilities are also enhanced at all levels. The average score of the test class was 8.8 points higher than that of the control class, and the average score was 10.78% higher. The results show that the application of new media in college music teaching is effective.

Early Domestication History of Asian Rice Revealed by Mutations and Genome-Wide Analysis of Gene Genealogies.

BACKGROUND: Asian rice (Oryza sativa L.) has been a model plant but its cultivation history is inadequately understood, and its origin still under debate. Several enigmas remain, including how this annual crop shifted its growth habit from its perennial ancestor, O. rufipogon, why genetic divergence between indica and japonica appears older than the history of human domestication, and why some domestication genes do not show signals of introgression between subgroups. Addressing these issues may benefit both basic research and rice breeding. RESULTS: Gene genealogy-based mutation (GGM) analysis shows that history of Asian rice is divided into two phases (Phase I and II) of about equal lengths. Mutations occurred earlier than the partition of indica and japonica to Os genome mark Phase-I period. We diagnosed 91 such mutations among 101 genes sampled across 12 chromosomes of Asian rice and its wild relatives. Positive selection, detected more at 5' regions than at coding regions of some of the genes, involved 22 loci (e.g., An-1, SH4, Rc, Hd3a, GL3.2, OsMYB3, OsDFR, and OsMYB15), which affected traits from easy harvesting, grain color, flowering time, productivity, to likely taste and tolerance. Phase-I mutations of OsMYB3, OsHd3a and OsDFR were experimentally tested and all caused enhanced functions of the genes in vivo. Phase-II period features separate cultivations, lineage-specific selection, and expanded domestication to more genes. Further genomic analysis, along with phenotypic comparisons, indicates that O. sativa is hybrid progeny of O. rufipogon and O. nivara, inherited slightly more genes of O. rufipogon. Congruently, modern alleles of the sampled genes are approximately 6% ancient, 38% uni-specific, 40% bi-specific (mixed), and 15% new after accumulating significant mutations. Results of sequencing surveys across modern cultivars/landraces indicate locus-specific usages of various alleles while confirming the associated mutations. CONCLUSIONS: Asian rice was initially domesticated as one crop and later separate selection mediated by human resulted in its major subgroups. This history and the hybrid origin well explain previous puzzles. Positive selection, particularly in 5' regions, was the major force underlying trait domestication. Locus-specific domestication can be characterized and the result may facilitate breeders in developing better rice varieties in future.

[Application of ultrasound-guided selective nerve branch blockage in lumbar spinal nerve posterior branch syndrome].

OBJECTIVE: To investigate the efficacy and safety of ultrasound-guided selective nerve branch blockage in the treatment of lumbar spinal nerve posterior branch syndrome. METHODS: A total of 40 patients with lumbar spinal nerve posterior branch syndrome treated by Pain Clinic from May 2017 to December 2018 were selected. According to the method used in locating site for nerve blockage, the patients were divided into ultrasound-guided group and anatomical positioning group, with 20 cases in each group. In anatomical positioning group, there were 7 males and 13 females, aged (63.42±7.71) years old, weighted (63.65±10.72) kg, numerical rating scale (NRS) was (6.61±1.52) scores, course of disease was (16.55±4.68) months. Pain sites:4 cases at L2,3, 8 cases at L3,4, 11 cases at L4,5, and 11 cases at L5S1. In ultrasound-guided group, there were 10 males and 10 females, aged (59.58±10.21) years old, weighted (60.61±13.81) kg, NRS was(6.84±2.43) scores, and course of disease was(13.70±5.98) months. Pain sites:6 cases at L2,3, 6 cases at L3,4, 9 cases at L4,5, and 13 cases at L5S1. Ultrasound-guided group used ultrasound-guided selective posteromedial branch and posterolateral branch nerve blockage, and the anatomical positioning group used anatomical localization method to block the posteromedial branch and posterolateral branch of lumbar spinal nerve. Each nerve branch was injected 3 ml of 0.125% ropivacaine. The number of treatment required and prone position time of each treatment were recorded, and the NRS scores of patients at the time points of immediately after the end of the treatment, the first week, the second week, the first month and the third month were evaluated. And adverse events such as local anesthetic allergy and poisoning, local puncture infection, total spinal anesthesia, dizziness, drowsiness, nausea, vomiting and other adverse reactions were observed. RESULTS: There were no statistically significant differences in gender, age, weight, NRS, course of disease and pain segment distribution between two groups (P>0.05). The number of treatment required in anatomical positioning group was significantly higher than that in ultrasound-guided group (P<0.000 1). During each treatment, the time in the prone position of the patients in anatomical positioning group was significantly lower than that in ultrasound guided group (P< 0.000 1). NRS scores immediately after the end of treatment, 1 week, 2 weeks, 1 month and 3 months, anatomical positioning group were 2.98 ±0.25, 3.04 ±0.38, 3.37 ±0.47, 3.42 ±0.85, 3.50 ±0.43, respectively, 2.94 ±0.31, 3.00 ±0.29, 3.21 ±0.68, 3.16 ± 0.94, 3.17±0.53 in ultrasound-guided group, and there was significant difference at 1 month and 3 months between two groups(P< 0.05). There were no adverse events such as local anesthetic allergy and poisoning, local puncture infection, and total spinal anesthesia, and no adverse reactions such as lethargy, nausea, and vomiting occurred in two groups. There were 6 cases of dizziness in anatomical positioning group and 12 cases in ultrasound guided group. The difference between two groups was statistically significant(P<0.05). CONCLUSION: Comparedwith anatomicalpositioning, ultrasound-guided selective nerve branch block for the treatment of posterior branch of the lumbar spinal cord syndrome can reduce the number of treatments and maintain a longer therapeutic effect, but it is also necessary to pay attention to the time of each treatment to avoid dizziness and other adverse reactions.

[Influence of different processed methods on 10 kinds of ginsensides in Panacis Quinquefolii Radix].

The influence on 10 kinds of ginsensides of different processed methods of Panacis Quinquefolii Radix was discussed. White Panacis Quinquefolii Radix (sliced and dried at -80 °C), red Panacis Quinquefolii Radix( steamed, sliced and dried at -80 °C) and commercial Radix Panacis Quinquefolii (dried by electric blast air) processed by different methods. HPLC-PDA-ESI- MS method was established before by our team. Ten kinds of ginsenosides of them were determined. The content of total ginsenosides were as follow: commercial Panacis Quinquefolii Radix > white Panacis Quinquefolii Radix > red Panacis Quinquefolii Radix. Compared with white Panacis Quinquefolii Radix, the content of Re, Rc, Rb3 and Rb2 of Red Radix Panacis Quinquefolii decreased but increased that of Rg,, Rb1. Both Rg2 and Rg, were not found in white Panacis Quinquefolii Radix and commercial Panacis Quinquefolii Radix by PDA detector, and low response in ESI-MS, while red Panacis Quinquefolii Radix was to the high content that of 0. 027% and 0.040 1%. The constituent of RA0 of red Panacis Quinquefolii Radix was higher than the other two. After Panacis Quinquefolii Radix processed, the kind and content of ginsensides were significantly changed. The constituent of some kinds of ginsensides was increased and some decreased. Rf was not found in all Panacis Quinquefolii Radix samples which were consistent with the former documents.

[Research on dynamic accumulation of nine ginsenosides and two pseudo-ginsenosides of Panax quinquefolium root of different growth years and harvest months in Canada].

OBJECTIVE: To research the dynamic accumulation of ginsenosides Rg1, Re, Rb1, Rg2, Rc, Rb2, Rb3, Rd, Rg3 and pseudo-ginsenosides F11,RT5, which was grown from one- to five-year-old and harvested from May to September of Panax quinquefolium root in Canada. METHODS: RP-HPLC-ELSD was adopted. The analysis was performed on a Dimaonsil C18 (250 mm x 4.6 mm, 5 µm) column, the mobile phase was acetonitrile (A)-deionized water (B) in gradient elution mode. The flow rate was 1.0 mL/min and column temperature was set at 30 °C. The carrier gas was nitrogen with flow rate of 2.8 L/min, the drift tube temperature was 100 °C, and the injection volume was 10 µL. The principle component analysis and cluster analysis of SPSS software (version 19.0) were conducted for the data analysis. RESULTS: The RP-HPLC-ELSD method for determining simultaneously nine ginsenosides and two pseudo-ginsenosides was established. The total content of ginsenosides of one- to two-year-old was low and increased from May to September in each year. That of three- to five-year-old declined from June to July and increased from August, and was the highest in September of three- to five-year-old samples. The total content from September of three-year-old samples was similar. The content of nine ginsenosides and two pseudo-ginsenosides was relatively high in all the samples except ginsenoside Rc, which wasn't determined in some samples. The ginsenosides Rd, Rb3 and pseudo-ginsenoside F11 were higher than others. The principle component analysis results showed that ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3, Rg3 and pseudo-ginsenosides F11,RT5 could be the characteristic ginsenosides of Panax quinquefolium root in Canada. The cluster analysis indicated that the chemical constituent and dynamic accumulation of ginsenosides of one- to two-year-old was similar except the sample of July of four-year-old and there was no obvious difference of three- to five-year-old. CONCLUSION: With the consideration of the content of ginsenoside, cultivation cost of four- to five-year-old, and risk of waterlogging and plant diseases, it is suggested that Panax quinquefolium root from September of three-year-old can be regarded as the same quality.

Characterization of tibetan medicine zuota by multiple techniques.

Zuota is regarded as the king of Tibetan medicine. However, due to the confidentiality of this precious medicine, the scientific characterization of Zuota is very scarce, which limits the pharmacology and biosafety studies of Zuota. Herein, we collected four different Zuota samples from Tibet, Qinghai, Gansu, and Sichuan and characterized them by multiple techniques. Our results showed that Zuota was mainly an inorganic mixture of HgS, sulfur, and graphite. Morphologically, Zuota samples were composed of nanoparticles, which further aggregated into microsized particles. Chemically, the majorities of Zuota were S and Hg (in the forms of HgS and pure sulfur). All samples contained pure sulfur with orthorhombic crystalline. Zuota from Qinghai province had different HgS crystalline, namely, hexagonal crystalline. The others were all face-centered cubic crystalline. Carbon in Zuota NPs was in the form of graphite. The implication to future studies of Zuota was discussed.