Whole genome and transcriptome reveal flavone accumulation in Scutellaria baicalensis roots.

Scutellaria baicalensis Georgi is a medicinal plant in the Lamiaceae family that contains high levels of 4'-deoxyflavone and other flavonoids in its roots. Therefore, it has strong potential as a plant resource for researching the biosynthesis of specific flavonoids. In this study, we report on a chromosome-level S. baicalensis genome assembled to nine chromosomes (376.81M) using PacBio, HiSeq XTen, and Hi-C assisted assembly. The assembly ratio was 99.22%, the contig N50 was 1.80 million bases, and the scaffold N50 was 40.57 million bases, with 31896 genes being annotated. Comparative genome analysis revealed that S. baicalensis and Salvia miltiorrhiza belonged to the same branch, and diverged 36.3 million years ago. Other typically correlated species were Boea hygrometrica and Sesamum indicum. We investigated the structural genes involved in flavonoid synthesis in combination with transcriptome sequencing analysis for different tissues (roots, stems, flowers, leaves) of purple, pink, and white flowers. The results revealed that S.baiF6H is involved in the accumulation of baicalein and was significantly increased in both purple roots vs. pink roots and white roots vs. pink roots. S.baiMYB gene family expression pattern analysis and co-expression network analysis revealed that S.baiMYB transcription factors primarily regulated the production of flavonoids in S. baicalensis. S.baiMYB serves as a major factor regulating flavonoid synthesis in the roots, where yeast one-hybrid assays revealed that these transcription factors could bind to the promoter regions of structural genes to control the accumulation of flavonoids. Genome and transcriptome sequencing, co-expression analysis, and yeast one-hybrid experiments provided valuable genetic resources for understanding flavonoid biosynthesis in S. baicalensis. These findings contribute to a better understanding of the accumulation of metabolites in Lamiaceae.

Serum Cystatin C Level Monitoring for Intervention Opportunity of CBP in Children with Severe Sepsis.

Objective: The aim of this study is to investigate the instruction value of the serum cystatin C (Cys C) level monitoring for intervention opportunity of continuous blood purification technology (CBP) in children with severe sepsis. Methods: 67 children with severe sepsis in the pediatric intensive care unit (PICU) with CBP treatment were retrospectively selected from May 2016 to April 2020. According to the time intervals between the time point of serum Cys C level began to increase (>15 mg/L) and the time point of CBP began, all children were divided into group A (<24 h, 29 cases), group B (24-48 h, 22 cases), and group C (>48 h, 16 cases). The children's general characteristics, vital signs, biochemical parameters, acute physiology and chronic health evaluation (APACHE II), and sequential organ failure assessment (SOFA) scores were evaluated. The influence factors of prognosis of children with severe sepsis were analyzed by multivariate regression analysis. Results: The intervals between the time point of PICU hospitalization and the time point of CBP began and the times of CBP in group A were significantly more than those in group B and C (P < 0.05). There was no statistically significant duration of CBP among three groups (P > 0.05). After follow-up of 28 d, there was no significant difference on the occurrence of coagulation disorders and hypovolemic shock induced by CBP among three groups (P > 0.05). However, the mortality of children in group A was lower than that in group C (P < 0.05). Children in group A had lower APACHE II scores, SOFA scores, serum K+, blood urea nitrogen (BUN), serum creatine (SCr), partial pressure of carbon dioxide (PCO2), and higher partial pressure of oxygen (PO2) than those of children in group C after CBP. (P < 0.05). SOFA scores ≥5 after CBP treatment and the time intervals between the time point of serum Cys C level began to increase (>15 mg/L) and the time point of CBP began ≥24 h were the independent influence factors on the prognosis by multivariate regression analysis. Conclusion: There are significant evidences that continuous blood purification technology within 24 h of serum Cys C level may better control the condition of children with severe sepsis.

Mechanism of Huangjing Qianshi Decoction in treatment of prediabetes based on network pharmacology and molecular docking / 中国中药杂志

This study analyzed the molecular mechanism of Huangjing Qianshi Decoction(HQD) in the treatment of prediabetes based on network pharmacology and molecular docking. The active components of HQD were identified and screened based on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP, http://Lsp.nwu.edu.cn/tcmsp.php) and then the targets of the components and the genes related to prediabetes were retrieved, followed by identifying the common targets of the decoction and the disease. The medicinal component-target network was constructed by Cytoscape to screen key components. The protein-protein interaction(PPI) network was established by STRING and hub genes were identified by Cytoscape-CytoNCA, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) of the hub genes with R-clusterProfi-ler. Thereby, the possible signaling pathways were predicted and the molecular mechanism was deduced. A total of 79 active components of HQD and 785 diabetes-related targets of the components were screened out. The hub genes mainly involved the GO terms of tricarboxylic acid cycle, peptide binding, amide binding, hydrolase activity, and kinase activity regulation, and the KEGG pathways of AGE-RAGE signaling pathway, TNF signaling pathway, AMPK signaling pathway, IL-17 signaling pathway, and insulin signaling pathway. Western blot result showed that HQD-containing serum significantly reduced the expression of AKT1, AGE, and RAGE proteins in insulin resistance model cells. HQD's treatment of prediabetes is characterized by multiple pathways, multiple targets, and multiple levels. The main mechanism is that the components zhonghualiaoine, baicalein, kaempferol, and luteolin act on AKT1 and inhibit the AGE-RAGE axis.

Pentahydroxy flavonoid isolated from Madhuca indica ameliorated adjuvant-induced arthritis via modulation of inflammatory pathways.

Rheumatoid arthritis (RA) is an autoimmune disease associated with advanced joint dysfunction. Madhuca indica J. F. Gmel, from the family Sapotaceae, is an Indian medicinal plant reported to have an array of pharmacological properties. The aim of present investigation was to determine the anti-arthritic potential of an isolated phytoconstituent from methanolic leaf extract of Madhuca indica (MI-ALC) against FCA-induced experimental arthritis. Polyarthritis was induced in female rats (strain: Wistar) via an intradermal injection of FCA (0.1 mL) into the tail. Polyarthritis developed after 32 days of FCA administration. Then rats were treated orally with an isolated phytoconstituent from MI-ALC at doses of 5, 10, and 20 mg/kg. Findings suggested that High-Performance Thin-Layer Chromatography, Fourier-Transform Infrared Spectroscopy, and Liquid Chromatography-Mass Spectrometry spectral analyses of the phytoconstituent isolated from MI-ALC confirmed the structure as 3,5,7,3',4'-Pentahydroxy flavone (i.e., QTN). Treatment with QTN (10 and 20 mg/kg) showed significant (p < 0.05) inhibition of increased joint diameter, paw volume, paw withdrawal threshold, and latency. The elevated synovial oxidative stress (Superoxide dismutase, reduced glutathione, and malondialdehyde) and protein levels of Tumor necrosis factor-α (TNF-α) and Interleukin (ILs) were markedly (p < 0.05) reduced by QTN. It also effectively (p < 0.05) ameliorated cyclooxygenase-2 (COX-2), Nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-kß) and its inhibitor-α (Ikßα), and ATP-activated P2 purinergic receptors (P2X7) protein expressions as determined by western blot analysis. In conclusion, QTN ameliorates FCA-induced hyperalgesia through modulation of elevated inflammatory release (NF-kß, Ikßα, P2X7, and COX-2), oxido-nitrosative stress, and pro-inflammatory cytokines (ILs and TNF-α) in experimental rats.

De Novo Assembly and Analysis of Polygonatum sibiricum Transcriptome and Identification of Genes Involved in Polysaccharide Biosynthesis.

Polygonatum sibiricum polysaccharides (PSPs) are used to improve immunity, alleviate dryness, promote the secretion of fluids, and quench thirst. However, the PSP biosynthetic pathway is largely unknown. Understanding the genetic background will help delineate that pathway at the molecular level so that researchers can develop better conservation strategies. After comparing the PSP contents among several different P. sibiricum germplasms, we selected two groups with the largest contrasts in contents and subjected them to HiSeq2500 transcriptome sequencing to identify the candidate genes involved in PSP biosynthesis. In all, 20 kinds of enzyme-encoding genes were related to PSP biosynthesis. The polysaccharide content was positively correlated with the expression patterns of ß-fructofuranosidase (sacA), fructokinase (scrK), UDP-glucose 4-epimerase (GALE), Mannose-1-phosphate guanylyltransferase (GMPP), and UDP-glucose 6-dehydrogenase (UGDH), but negatively correlated with the expression of Hexokinase (HK). Through qRT-PCR validation and comprehensive analysis, we determined that sacA, HK, and GMPP are key genes for enzymes within the PSP metabolic pathway in P. sibiricum. Our results provide a public transcriptome dataset for this species and an outline of pathways for the production of polysaccharides in medicinal plants. They also present more information about the PSP biosynthesis pathway at the molecular level in P. sibiricum and lay the foundation for subsequent research of gene functions.

[Inhibition effects and mechanisms of the entophytic fungus Trichoderma harzianum LH-7 from Aloe barbadensis].

Inhibition spectrum and antagonistic mechanism of an endophytic fungus Trichoderma harzianum LH-7, isolated from wild medicinal plant Aloe barbadensis, were investigated by in vitro culture methods against 9 kinds of plant pathogens. The results showed that nutrient competition and hyper-parasitism were the two primarily antagonist approaches that strain LH-7 adopted to inhibit the tested plant pathogens with a significant inhibition rate of 62.4%-88.4%. Moreover, the active compound from metabolites of LH-7 could cause pathogen mycelial deformities, cell wall rupture and conidial malformation, leading to the effective inhibition on pathogens growth and reproduction.

[Effects of electroacupuncture on the proprioception of athletes with functional ankle instability].

OBJECTIVE: To observe the efficacy differences between electroacupuncture and physiotherapy on the proprioception of athletes with functional ankle instability (FAD. METHODS: Fifty athletes with FAI were randomly divided into an electroacupuncture group and a physiotherapy group. The electroacupuncture group was treated with electroacupuncture at Jiexi (ST 41), Kunlun (BL 60), Qiuxu (GB 40) and Ashi acupoints, and the physiotherapy group was treated with low frequency electrical stimulation and infrared radiation at medial malleolus and lateral malleolus, thrice each week for consecutive 8 weeks. The Joint Position Sense: Active (JPSA), Joint Position Sense: Passive (JPSP) and Kinaesthesia (KT) were assessed at the ankle by use of Biodex System-III isokinetic dynamometer. RESULTS: The JPSA of 11.090 +/- 3.1 degrees and the JPSP of 9.67 degrees +/- 2.8 degrees before the treatment reduced to 9.14 degrees +/- 4.0 degrees and 6.89 degrees +/- 3.3 degrees, respectively, after the treatment in the electroacupuncture group, with significant differences in JPSA and JPSP (both P < 0.05), compared with those in the physiotherary group, there were significant differences (both P < 0.05) and with no significant difference in KT (P > 0.05). There was no significant differences in the indices of JPSA, JPSP and KT in the physiotherapy group after 8 weeks than those before treatment (all P > 0.05). CONCLUSION: Electroacupuncture can effectively improve the proprioception of athletes with FAI and achieves a superior efficacy as compared with the conventional physiotherapy.

Observation on therapeutic effect of cervical spondylosis of vertebral artery type treated with both acupuncture and mild moxibustion / 中国针灸

<p><b>OBJECTIVE</b>To observe the clinical therapeutic effects on cervical spondylosis of vertebral artery type treated with both acupuncture and mild moxibustion and investigate the mechanism of action.</p><p><b>METHODS</b>Sixty cases were randomly divided into the combined therapy group of acupuncture and mild moxibustion (combined therapy group) and the acupuncture group, 30 cases for each group. In combined therapy group, Baihui (GV 20), Fengchi (GB 20) and cervical Jiaji (EX-B 2) were punctured, and Baihui(GV 20), Dazhui (GV 14), Mingmen (GV 4), Shenque (CV 8) and Guanyuan (CV 4) were treated with mild moxibustion; in acupuncture group, acupuncture was simply applied, the acupoints selection and manipulation were the same as those in the combined therapy group. The average velocity meter and the resistive index of vertebral and basal arteries were observed before and after treatment by TCD examination, the therapeutic effects and clinical symptom-sign scores were observed too.</p><p><b>RESULTS</b>The total effective rate was 93.3% (28/30) in combined therapy group, and 73.3% (22/30) in acupuncture group. The therapeutic effect in combined therapy group was superior to that in acupuncture group (P < 0.05). The improvements of symptom-sign scores in two groups after treatment were more significant than that before treatment, and the score in the combined therapy group was superior to that in acupuncture group (P < 0.05). The average velocity meter and the resistive index of vertebral and basal arteries after treatment were improved in both groups, in which, the improvement in combined therapy group was more apparent (all P < 0.05).</p><p><b>CONCLUSION</b>The therapeutic effects on cervical spondylosis of vertebral artery type treated with both acupuncture and mild moxibustion are preferable; by this therapy, the average velocity meter of vertebral and basal arteries are accelerated and the resistive index is decreased.</p>