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Abstract Angiotensin II (AngII) causes endothelial dysfunction. Eucommia ulmoides extract (EUE) is documented to manipulate AngII, but its impact on cardiac microvascular endothelial cell (CMVEC) function remains unknown. This study determines the effects of EUE on AngII-treated CMVECs. CMVECs were treated with different concentrations of AngII or EUE alone and/or the p53 protein activator, WR-1065, before AngII treatment, followed by examinations of the apoptotic, migratory, proliferative, and angiogenic capacities and nitric oxide (NO), p53, von Willebrand factor (vWF), endothelin (ET)-1, endothelial NO synthase (eNOS), manganese superoxide dismutase (MnSOD), hypoxia-inducible factor (HIF)-1α, and vascular endothelial growth factor (VEGF) levels. AngII induced CMVEC dysfunction in a concentration-dependent manner. EUE enhanced the proliferative, migratory, and angiogenic capacities and NO, MnSOD, and eNOS levels but repressed apoptosis and vWF and ET-1 levels in AngII-induced dysfunctional CMVECs. Moreover, AngII increased p53 mRNA levels, p-p53 levels in the nucleus, and p53 protein levels in the cytoplasm and diminishes HIF-1α and VEGF levels in CMVECs; however, these effects were counteracted by EUE treatment. Moreover, WR-1065 abrogated the mitigating effects of EUE on AngII-induced CMVEC dysfunction by activating p53 and decreasing HIF-1α and VEGF expression. In conclusion, EUE attenuates AngII-induced CMVEC dysfunction by upregulating HIF-1α and VEGF levels via p53 inactivation

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Eucommia ulmoides Oliver, a single extant species of Eucommiaceae, is an endemic dioecious tree in China. The natural resources of E. ulmoides have rapidly declined in recent years because of the over-collection of its cortex. To design a suitable protection strategy, it is necessary to develop a set of molecular markers to investigate genetic diversity and population structure of E. ulmoides. Pyrosequencing of an enriched microsatellite library by Roche 454 FLX+ platform was used to isolate simple sequence repeats (SSRs) for E. ulmoides. A total of 1568 SSRs that contained enough flanking sequences for primer pair design were identified from 45,236 raw sequence reads. One hundred SSRs were randomly selected to design primer pairs and polymerase chain reaction was performed. Among these 100 tested primer pairs, 16 were polymorphic across 18 individuals from three E. ulmoides populations. The number of alleles ranged from 3 to 8, with an average of 5.1. The expected heterozygosity ranged from 0.110 to 0.830, with an average of 0.648, and the observed heterozygosity ranged from 0.111 to 0.833, with an average of 0.524. The inbreeding coefficient ranged from -0.349 to 0.547. This set of microsatellite markers could be valuable for landscape genetic structure assessment and molecular marker-assisted breeding in E. ulmoides.

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Eucommia ulmoides Oliver, the only extant species of Eucommiaceae, is a second-category state-protected endangered plant in China. Evaluation of genetic diversity among some intraspecific hybrid populations of E. ulmoides Oliver is vital for breeding programs and further conservation of this rare species. We studied the genetic diversity of 130 accessions from 13 E. ulmoides intraspecific hybrid populations using inter-simple sequence related (ISSR) and sequence-related amplified polymorphism (SRAP) markers. Of the 100 ISSR primers and 100 SRAP primer combinations screened, eight ISSRs and eight SRAPs were used to evaluate the level of polymorphism and discriminating capacity. A total number of 65 bands were amplified using eight ISSR primers, in which 50 bands (76.9%) were polymorphic, with an average of 8.1 polymorphic fragments per primer. Alternatively, another 244 bands were observed using eight SRAP primer combinations, and 163 (66.8%) of them were polymorphic, with an average of 30.5 polymorphic fragments per primer. The unweighted pair-group method (UPGMA) analysis showed that these 13 populations could be classified into three groups by the ISSR marker and two groups by the SRAP marker. Principal coordinate analysis using SRAP was completely identical to the UPGMA-based clustering, although this was partly confirmed by the results of UPGMA cluster analysis using the ISSR marker. This study provides insights into the genetic background of E. ulmoides intraspecific hybrids. The progenies of the variations "Huazhong-3", "big fruit", "Yanci", and "smooth bark" present high genetic diversity and offer great potential for E. ulmoides breeding and conservation.

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Eucommia ulmoides Oliver, one of the tertiary relict species found only in China, is the only extant species of Eucommiaceae. Using inter-simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers, we studied the genetic diversity and population genetic structure of 187 accessions from 17 E. ulmoides populations throughout its main distribution in China. A total of 65 bands were amplified using eight ISSR primers, of which 50 bands (76.9%) were polymorphic. Meanwhile, another 244 bands were observed using eight SRAP primer combinations and 163 (66.8%) of these were polymorphic. The analysis of molecular variation (AMOVA) indicated that 88.8 and 92.4% of the total variation resided within populations based on ISSR and SRAP analysis, respectively. Moreover, we found that the E. ulmoides populations were clustered into six distinct groups using ISSR and SRAP markers via the unweighted pair-group method (UPGMA). Furthermore, STRUCTURE analysis showed that these 17 populations could be classified into four groups using ISSR markers, but only two groups using SRAP markers. No significant relevancy was observed between genetic and geographic distances among the sampled populations. The results of this study support the view that exchange of seeds among local farmers plays an important role in shaping the present genetic distribution pattern. "Core collection" is suggested for genetic diversity conservation of E. ulmoides in China.

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Eucommia ulmoides is cultivated for the production of eucommia rubber and Chinese herbal drugs. Molecular breeding methods, such as marker-assisted selection (MAS), have the potential to improve the efficiency of E. ulmoides breeding. Quantitative trait locus (QTL) analysis was applied to identify marker-trait associations for E. ulmoides using an F1 mapping population of 152 individuals derived from a cross between the wild genotype Xiaoye and the cultivar Qinzhong No. 1. A total of 78 QTLs were identified for 12 leaf traits involving morphology, yield, and secondary metabolites. Phenotypic variance explained by individual QTLs ranged from 10.4 to 53.3%. Fifteen QTL clusters, each harboring loci controlling at least two leaf traits, were detected across nine linkage groups. Co-location of these QTLs may be due to pleiotropy or linkage. Three main QTL regions for secondary metabolites were consistently identified each year. QTL information from this study furthers our understanding of the genetic architecture of these economically important traits and of MAS in E. ulmoides breeding.

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Sequence-related amplified polymorphism (SRAP), amplified fragment length polymorphism (AFLP), and inter-simple sequence repeat (ISSR) markers were used to estimate the genetic diversity and relationships among Eucommia ulmoides cultivars in China. A total of 240, 192, and 150 DNA fragments were detected by 10 SRAP primer combinations, 10 AFLP primer combinations, and 10 ISSR primers, among which 89.2, 65.1, and 88.0% of the fragments were polymorphic, respectively. Cluster analysis revealed that Qinzhong No. 3, Xiaoyeci, Qinzhong No. 1, and Qinzhong No. 2 formed independent clusters. The other 15 cultivars exhibited two clusters. The results of this study will help in the selection of parents for both genome mapping and crossbreeding purposes.

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Progeny performances, variations and combining abilities for growth traits were evaluated in a factorial mating design of Eucommia ulmoides. Three marker systems, sequence-related amplified polymorphism, amplified fragment length polymorphism, and inter-simple sequence repeat, were used to determine genetic distances between parents. Correlations of genetic distances with progeny performances, within-family coefficients of variation and specific-combining abilities were established for height and basal diameter traits. Significant positive correlations were found between progeny performances of growth traits and genetic distances of parents based on sequence-related amplified polymorphism markers or a combination of all 3 marker systems. This revealed that crosses between genetically distant parents produced progenies with excellent growth performances. The lack of correlations between parental genetic distances and within-family coefficients of variation or specific-combining abilities suggested that these characteristics were unpredictable. The results of this study represent a potential criterion to predict progeny performances and choose parents in the breeding program.

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En los últimos años se ha incrementado el interés por el uso de terapias alternativas para la curación de heridas. La miel de abeja es una antigua medicina natural que ha sido utilizada en el tratamiento de heridas y úlceras, mientras la vitamina C mejora la barrera epidérmica y reduce la contracción de heridas. Este estudio pretende avalar el tratamiento con miel asociada a vitamina C oral como curación alternativa y eficiente en quemaduras tipo AB-B. Se utilizaron 12 cuyes (Cavia porcellus). Los animales se distribuyeron en grupos A (sin Vitamina C) y B (con Vitamina C). Bajo efectos anestésicos se provocó una quemadura de 1.7 mm de diámetro en la región dorsal derecha, que fue tratada con curaciones con miel. Al mismo tiempo, se provocó una quemadura de similares características en la región dorsal izquierda, tratada con suero Ringer, siendo utilizada como control. De ambas lesiones se obtuvieron biopsias a los 6, 10 y 14 días post injuria. Nuestros resultados muestran que la formación de tejido granular, activación de fibroblastos, y reepitelización es más rápida en el grupo B, donde se observó un tejido vascularizado y fibras colágenas organizadas en el día 14 post injuria, similar al control (biopsia de animal sano). Estos datos confirman que la utilización de la miel de ulmo asociada a Vitamina C oral mejora la regenaración de este tipo de heridas, logrando una cicatrización efectiva, rápida y de buena calidad.

In recent years there has been increasing interest in the use of alternative therapies for healing wounds. Bee honey is an ancient natural medicine that has been used in the treatment of wounds and ulcers, while Vitamin C improves the epidermal barrier and reduces wound contraction. The aim of this study is to support treatment with honey associated with oral vitamin C as an alternative and efficient healing treatment, in type AB-B burns. For this study, twelve guinea pigs (Cavia porcellus) were used. The animals were distributed in groups A (without Vitamin C) and B (with Vitamin C). While under anesthetic effect burn of 1.7 mm in diameter was caused in the right dorsal region, subsequent healing was treated with honey. At the same time a burn of similar characteristics was caused in the left dorsal region, treated with Ringer serum being used as control. Biopsies were obtained from both lesions at days 6, 10 and 14 post injury. Results showed granular tissue formation, fibroblast activation, and reepitalization is more rapid in group B, where vascularized tissue and organized collagen fibers were observed at day 14 post injury similar to control (biopsy in healthy animal). These results confirm that the use of Ulmo honey associated with oral Vitamin C improves regeneration in this type of wound, achieving effective, rapid and good quality healing.

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1. Changes in the cardiovascular parameters of Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were examined following a 4-week diet of either Brazilian propolis or Eucommia uloides OLIVER (tochu). 2. A 4-week diet of propolis or tochu resulted in significant reductions in systolic blood pressure in SHR but had no effect on WKY. Experiments using aorta isolated from animals fed a diet of propolis or tochu revealed increased acetylcholine-induced relaxation in SHR and no change in acetylcholine-induced relaxation in WKY. Sodium nitroprusside-induced relaxation was unaffected by propolis or tochu in both animal groups. 3. These results suggest that propolis and tochu produce an antihypertensive effect that may be mediated by potentiation of acetylcholine-induced vasodilatation.

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