Resolving complicated relationships of the Panax bipinnatifidus complex in southwestern China by RAD-seq data.

The P. binpinnatifidus complex included most of the Panax species distributed in Sino-Himalaya regions except for P. pseudoginseng, P. stipuleanatus and P. notoginseng. However, the delimitation and identification of these taxa within the species complex are very difficult due to the existence of morphological intermediates, and their evolutionary relationships remain unresolved despite several studies have been carried out based on traditional DNA markers. The taxonomic uncertainty hinders the identification, conservation and exploration of these wild populations of Panax. To study this species complex, we employed ddRAD-seq data of these taxa from 18 different localities of southwestern China, using two RAD analysis pipelines, STACKS and pyRAD. Based on the results of phylogenetic analysis, the species complex was divided into four clades with high supports, which largely agreed with morphologically described species. Two clades, corresponding to P. vietnamensis and P. zingiberensis, respectively, were sister groups, indicating that these two species had a closer genetic relationship; the third clade was consisted of samples with bamboo-like rhizomes named as P. wangianus clade, and the fourth one with moniliform rhizomes was named as P. bipinnatifidus clade. The population genetic structure analysis and D-statistics test showed the localized admixture among these species, which indicated that introgression had occurred among the related lineages continuously distributed in southeastern Yunnan and adjacent regions.

MicroRNA-381 regulates the occurrence and immune responses of coronary atherosclerosis via cyclooxygenase-2.

The present study aimed to measure the levels of microRNA-381 (miR-381) in the plaque tissues, peripheral blood mononuclear cells (PBMCs) and serum of patients with coronary atherosclerosis. In addition, the regulatory mechanisms of miR-381 and cyclooxygenase (COX)-2 in coronary atherosclerosis were investigated. A total of 36 patients with coronary atherosclerosis who received coronary endarterectomy at Linyi People's Hospital and Junan Hospital of Traditional Chinese Medicine (Linyi, China) between January 2013 and June 2016 were enrolled into the present study, while 39 healthy subjects were included as the control group. Peripheral blood was collected form all patients and healthy subjects. Plaque tissues were resected from patients with coronary atherosclerosis and adjacent artery intimal tissues were resected as the control tissues. Using quantitative polymerase chain reaction, the levels of miR-381 and COX-2 mRNA in the plaque tissues, PBMCs and serum were determined. In addition, COX-2 protein expression in the plaque tissues and PBMCs was measured by western blotting, while enzyme-linked immunosorbent assay was utilized to examine the protein content in the serum. To identify the direct interaction between miR-381 and COX-2 mRNA, dual-luciferase reporter assay was also conducted. The levels of COX-2 mRNA and protein in the plaque tissues, PBMCs and serum of patients with coronary atherosclerosis were significantly elevated compared with those in the corresponding control groups. However, the expression of miR-381 was significantly reduced in the coronary atherosclerosis patients. Dual-luciferase reporter assay revealed that miR-381 was able to directly target the 3'-untranslated region of COX-2 mRNA to regulate the expression of COX-2. Therefore, the present study demonstrated that enhanced levels of COX-2 expression in patients with coronary atherosclerosis are associated with the downregulation of miR-381 expression, while miR-381 may regulate the occurrence and immune responses of coronary atherosclerosis via COX-2.

Application of RAD Sequencing for Evaluating the Genetic Diversity of Domesticated Panax notoginseng (Araliaceae).

Panax notoginseng, a traditional Chinese medicinal plant, has been cultivated and domesticated for approximately 400 years, mainly in Yunnan and Guangxi, two provinces in southwest China. This species was named according to cultivated rather than wild individuals, and no wild populations had been found until now. The genetic resources available on farms are important for both breeding practices and resource conservation. In the present study, the recently developed technology RADseq, which is based on next-generation sequencing, was used to analyze the genetic variation and differentiation of P. notoginseng. The nucleotide diversity and heterozygosity results indicated that P. notoginseng had low genetic diversity at both the species and population levels. Almost no genetic differentiation has been detected, and all populations were genetically similar due to strong gene flow and insufficient splitting time. Although the genetic diversity of P. notoginseng was low at both species and population levels, several traditional plantations had relatively high genetic diversity, as revealed by the He and π values and by the private allele numbers. These valuable genetic resources should be protected as soon as possible to facilitate future breeding projects. The possible geographical origin of Sanqi domestication was discussed based on the results of the genetic diversity analysis.