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Long non‐coding RNAs (lncRNAs) have been identified as playing critical roles in multiple diseases. However, little is known regarding their roles and mechanisms in post‐stroke angiogenesis. Our studies focused on deciphering the functional roles and the underlying mechanisms of the lncRNA metastasis‐associated lung adenocarcinoma transcript 1 (MALAT1) in the process of angiogenesis following oxygen‐glucose deprivation/reoxygenation (OGD/R). We characterized the up‐regulation of MALAT1 expression in the process of angiogenesis after hypoxic injury in vivo and in vitro. We further showed that compared with the empty vector, MALAT1 knockdown had significantly reduced the capacity for angiogenesis, which was measured by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium (MTT), scratching, cell cycle and immunofluorescent staining. Thus, our findings suggest that MALAT1 may mediate proangiogenic function in OGD/R. To further explore the potential mechanisms, we used lentiviruses expressing shMALAT1 and empty vector; the results revealed that shMALAT1 reduced the expression of 15‐lipoxygenase 1 (15‐LOX1), vascular endothelial growth factor (VEGF) and the phosphorylation of signal transducers and activators of transcription 3 (pSTAT3). Taken together, our results are the first to propose that MALAT1 may regulate angiogenesis through the 15‐LOX1/STAT3 signalling pathway, and they may provide a critical target for the treatment of hypoxic injury and an avenue for therapeutic angiogenesis.

Background/aims With advances in RNA-sequencing (RNA-seq), exploring the expression and transcripts of different classes of genes are now possible. Our purpose was to analyse to the long non-coding RNAs (lncRNAs) in mouse brain microvascular endothelial cells (bEnd.3) after oxygen-glucose deprivation/re-oxygenation (OGD/R). Methods RNA-seq was employed to explore the expression of lncRNAs, and quantitative real-time PCR (qRT-PCR) was used to identify the results of RNA-seq. Furthermore, the biological functions of the lncRNAs were identified by cell viability, wound healing, transwell, tube formation and immunofluorescent staining assays. Finally, the molecular mechanisms involving the differentially expressed lncRNAs were further explored by bioinformatics and Western blotting (WB). Results In total, 2710 lncRNAs were found, 33 of which were significantly differentially expressed, with 18 upregulated lncRNAs and 15 downregulated lncRNAs in brain microvascular endothelial cells following OGD/R. Among the dysregulated genes, G protein-coupled receptor 137b-pseudogene (Gpr137b-ps), predicted gene 32856 (Gm32856), small nucleolar RNA host gene 17 (snhg17), chaperonin containing Tcp1 and subunit 6a (Cct6a) were significantly upregulated lncRNAs; this finding was further validated using qRT-PCR. Moreover, Gene Ontology (GO) and Kyoto Encyclopaedia of Genes (KEGG) pathway analyses were employed to decipher the potential target genes and signaling pathways of the differentially expressed lncRNAs. Finally, we selected pseudogene-expressed lncRNA Gpr137b-ps as a candidate gene, and report for the first time that pseudogenes can mediate angiogenesis and their potential target genes, namely, 15-lipoxygenase1 (15-LOX1), Signal Transducer and Activator of Transcription 3 (S TAT3) and vascular endothelial growth factor (VEGF). Conclusion Therefore, our study revealed that Gpr137b-ps plays critical roles in the process of angiogenesis, suggesting avenues for the development of future therapeutic strategies that contribute to promoting angiogenesis following I/R.

Three new isolates (

To investigate the influence of storage period and grading standards on the quality of Prunella vulgaris in order to provide a theoretical basis for standardized cultivation.The spices of P. vulgaris in different storage period (including current year, first-year and second-year) were collected from Lujiang base in Anhui Province. The samples were randomized, with the methods of morphologic characteristics and commercial size on P. vulgaris, the samples were graded into three grades; according to the methods from the Pharmacopoeia of the People's Republic of China, water-soluble extracts, ethanol-soluble extracts, total ash and acid insoluble ash were measured, respectively, utilizing the UV spectrophotometry to detection the contents of total flavonoids and polysaccharides, the levels of ursolic acid, oleanolic acid and rosmarinic acid were adopted by HPLC.The total ash and acid insoluble ash of P. vulgaris showed significantly increase during prolonged storage; however, the content of water-soluble extracts, ethanol-soluble, total flavonoids, polysaccharides, ursolic acid, oleanolic acid,rosmarinic acid showed significantly decrease during prolonged storage. The spica of P. vulgaris exhibited no rules differences between the quality and grading standards, The second-degree belongs to high content of total flavonoids; the high content of polysaccharides was first-degree; the total ash and acid insoluble ash of spicas exhibited gradually decreased from first-degree to third-grade, and the content of water-soluble extracts, ethanol-soluble extracts,ursolic acid,oleanolic acid, rosmarinic acid showed gradually increased from first-degree to third-grade.The current year was a suitable storage time of spicas from P. vulgaris could be selected; and the main bioactive components highest level of P. vulgaris spicas was third-degree, second-grade next, first-degree minimum.

Background The mechanism by which vascular regeneration declines with aging is not fully understood. An interaction between integrin and vascular endothelial growth factor receptor-2 (VEGFR-2) plays a substantial role in angiogenesis. Here, we investigated whether aging impairs this interaction in endothelial progenitor cells (EPCs) under hypoxia. Methods and results Aging reduced the blood flow and vessel density in ischemic muscles in mice. Levels of phosphorylated Src (p-Src), p-β 3 , and p-VEGFR-2 in acute ischemia were reduced in the muscles of aged mice compared to young mice. The hypoxia-inducible factor (HIF)-1α stabilizer deferoxamine improved the age-related impairment of angiogenesis, but this effect was diminished by LY290004, an inhibitor of phosphatidylinositol 3-kinase. Deferoxamine improved the reduction in chronic ischemia-induced β 3 -integrin and VEGFR-2 phosphorylation in the muscles of aged mice; this effect was also diminished by LY290004. In EPCs, we identified the molecular requirements for VEGF-mediated β 3 -integrin and VEGFR-2 cross-activation in vitronectin-induced cell adhesion under acute hypoxia. We demonstrated that c-Src controlled the adhesion- and VEGF-induced β 3 tyrosine phosphorylation in hypoxia. Aging enhanced the hypoxia-induced EPC apoptosis and impaired several c-Src-related VEGF-induced receptor events, including β 3 tyrosine activation, ligand binding, cell adhesion, and tubulogenesis in cultured EPCs of animals and those of humans. Conclusions These data suggest that the aging-related decline in angiogenic action in response to ischemia is mediated by the impairment of cross-activation between β 3 and VEGFR-2 in EPCs, which is partially associated with decreased HIF-1α stability.

OBJECTIVE The accumulation of dry matter, nitrogen, phosphorus and potassium in Prunella vulgaris and their relationships under field condition were studied, in order to provide a basis for the rational fertilization. METHOD Through the dynamic sampling of P. vulgaris in different growing stages, and the dry weight of plant, the amount of dry matter and the content of N, P and K within each organ of the plant were measured. RESULT The dry weight of P. vulgaris linearly increased with the increase of nitrogen, phosphorus and potassium accumulation, there existed a significant correlation at whole growth period. The maximum stage accumulation rate of dry matter from mid-April to the beginning of May, reached 63.71%. The amount of dry matter was mainly distribution in leaves from late March to mid-April. The distribution ratio in stem was the highest from beginning of May to late May. Until mid-June, the distribution ratio of dry matter in spicas was higher than that in other organs. The absorption amounts of N, P and K were lower before mid-April, and the nutrient element was concentrated in plant leaves. From late April to beginning of May, the N, P and K accumulation reached maximum of total absorptive capacity, which accounted for 72.35%, 59.82%, 87.41%, respectively, and the mineral elements were concentrated distribution of plant stems. When transferred to reproductive stage, both accumulation amount and distribution rate of nutrient elements in spicas increased rapidly, which reached the maximum until mid-June. From elongation stages to quaring period was the period of maximal accumulation of dry matter and showed the maximal efficiency of N, P and K nutrition. CONCLUSION The time from elongation stages to quaring period is the most crucial period in the growth development stage of P. vulgaris, and should be paid more attention to the supply of mineral nutrition and moisture of plant to promote spicas formation and dry matter accumulation in this stage. The average N, P2O5 and K2O absorption amount per 100 kg dry matter of spicas were 1.62, 0.36, 2.88 kg respectively, and the ratio of N-P2O5,-K2O was 1 : 0.22: 1.77.

Objective The contents of mineral elements in soil and each organ from cultivated and wild Prunella vulgaris were determined in order to provide a theoretical basis for exploitation and protection of wild resources and GAP study. Method The contents of mineral elements (P, K, Ca, Mg, Na, Fe, Cu, Zn, Mn, Al) in soil and each organ from cultivated and wild P. vulgaris were determined by ICP-AES, and the results were analyzed by correlation analysis. Result The contents of mineral elements in soil and different organs of P. vulgaris were significant different between cultivated and wild species; the contents of P and Cu ranked in the order of spica > leaf > stem, the order of Mg, Ca, Zn, Fe, Mn, Al from high to low was leaf > spica > stem, the concentrations of K, Na were mainly distributed in leaf and stem, less in spica. A distinctly positive correlation between soil and spica for Fe content, and the relationship between soil and leaf, stem were significant positive correlation, the concentrations of Zn in spica, leaf, stem were positively related to soil, K contents in soil and stem were significant positive correlation. The different organs of cultivated and wild P. vulgaris had a strong tendency to accumulate P. Conclusion There was no direct relationship between the concentrations of mineral elements in P. vulgaris, and their corresponding soil P. vulgaris adjusts the concentration of mineral elements mainly by active absorption.

OBJECTIVE To study the effects of nitrogen (N), phosphorus (P2O5), potassium (K2O) and organic compound fertilizer (OF) on spicas biomass and yield of Prunella vulgaris in order to provide a theoretical basis for good agriculture practice (GAP). METHOD Field plot experiments with a 4-variable quadratic orthogonal rotation combination design were adopted, and a function model of spicas yield in P. vulgaris was established. RESULT The model analysis showed that spicas yields of P. vulgaris were significantly influenced under the N, P2O5 and K2O applications, among there factors, N had the greatest effects on spicas yield of P. vulgaris, the next was P2O5 and K2O, OF had the smallest effects on spicas yield. At the lower fertility levels, the spicas yields of P. vulgaris were improved along with the increasing of N, P2O5, K2O and OF applications, but the spicas yields decreased when N, P2O5, K2O and OF were applied too much. CONCLUSION In this experiment condition, the spicas yields of P. vulgaris could reach to 722 kg x hm(-2) when the fertilizer rates were N 303. 9-335.1 kg x hm(-2), P2O5 432.5-500.6 kg x hm(-2), K2O 206.6-240.2 kg x hm(-2), OF 2 312.5-2 687.5 kg x hm(-2).

OBJECTIVE To analyze the dynamic change of chemical constituents in Changium smyrnioides at different ages and give a reference for standard cultivation. METHOD The contents of polysaccharides, mannitol and choline in the roots were determined by colorimetry. HPLC was used to analyze water-soluble components in the roots. GC-MS was used to analyze volatile oil components in the leaves. Similarity coefficient of water-soluble components was calculated by included angle cosine method. RESULT There was significant difference in content of the analyzed chemical constituents except polysaccharide. Contents of mannitol, physiological ash and polysaccharide in the adult plant were higher than those in the young plant. On the contrast, content of water-soluble components and choline in the young plant was higher than that in the adult plant. With the increase of the growth years, water-soluble components in the roots and volatile oil components in the leaves were gradually concentrated. CONCLUSION The growth time significantly affects the chemical constituents in Ch. smyrnioides, the reasonable harvest time is from the second to the forth year after transplanting.

To analyze the distribution of chemical constituents in different parts of Changium smyrnioides.The content of polysaccharides, mannitol and choline was determined by colorimetry and the water-soluble components was determined by HPLC. similarity coefficient of water-soluble components was calculated by the included angle cosine method.The Contents of chemical constituents were significantly different between the leaves and the roots. The contents of choline and polysaccharides in the roots was higher than those in the leaves, but the contents of water-soluble extract and physiological ash in the leaves was higher than those in the roots. Contents of chemical constituents were also significantly different at different segments in the root, which decreased from the upper to the lower parts, increased from the outside to the inside parts except physiological ash, and the difference in horizontal segments was higher than that in vertical segments. Total peak area of water-soluble components in the leaves was 2.81 times higher than that in the roots, and gradually increased from the outside to the inside parts.Total content of chemical constituents in the leaves was significantly higher than that in the roots. The quality of medicinal used parts decreased gradually from the inside to the outside parts and from the upper to the lower parts in the root of Ch. smyrnioides.

OBJECTIVE To investigate the herbal medicine of Prunella vulgaris by textual researches, and provide a theoretical basis for clinical medication, exploitation and protection of wild P. vulgaris resources. METHOD Textual research on medicinal works of past dynasties and field work were adopted. RESULT The natural distributions of P. vulgaris were concentrated distribution in Sichuan province, Huaihe river basin, and Middle-Lower Yangtze river valley in Chinese history. The indications of P. vulgaris in ancient and modern times were basically identical. While there were difference between the medicinal parts, harvest period and processing methods existed difference between ancient and modern. CONCLUSION Three periods that whole grass of P. vulgaris as medicinal parts (from late Ming dynasty to late Qing dynasty and early stage of Republic of China), both whole grass and spicas as medicinal parts (from mid-term Republic of China to 1963), and the semi-maturity or maturity of spicas as medicinal parts (from 1963 to today) existed. The processing method for medicinal parts of P. vulgaris adopted sun drying and shady drying in ancient China, but only the sun drying was only used in modern times.

To study the correlation among the morphological characteristics and the spica yield of various germplasm of Prunella vulgaris.The various P. vulgaris germplasm from all over the country were investigated by analysis of correlation analysis, path analysis and principle component analysis in a randomized block experiment.The 6 morphological characteristics were greatly different from each other in various germplasm. The spica yield per plant had a very significantly positive correlation with the number of spica per plant and fresh leaves weight per plant, meanwhile the correlation between the spica yield per plant and spica length was very significantly. Three principal components which accounted for 87.533% of total variance were extracted from the principal component analysis.The strong growth potential, the number of spica per plant and spica length were main factors for the selection of high yield breeding of P. vulgaris.

To investigate the insulin-like growth factor-I (IGF-I) gene and polypeptide expression in cultured rat osteoblast (ROB) and the role of IGF-I in mediating the cell-to-cell communication by mimicking the pharmacokinetics of parathyroid hormone (PTH).The ROB was cultured with three kinds of treatment: (1) Control (Ctr), the cells were cultured without PTH during the first 6 hours and the subsequent 42 hours in a 48-hour cycle; (2) Intermittent exposure to PTH (Itm), the cells were cultured with PTH during the first 6 hours, but without PTH in the subsequent 42 hours; and (3) Continuous exposure to PTH (Ctu), the cells were cultured with PTH during the first 6 hours and the subsequent 42 hours.The bone-forming activities of ROB were increased in Itm and inhibited in Ctu. The IGF-I mRNA content in Itm cells was elevated only during the first 6 hours and that in Ctu cells was elevated at any time during an incubation cycle. The free IGF-I concentration in the medium of Itm cells was generally higher and that of the Ctu cells was generally lower compared with those of the Ctr cells. The IGF-I antibody significantly reduced the alkaline phosphatase activity within the cells of Ctr and Itm.PTH rapidly and constantly stimulates the IGF-I gene transcription of osteoblast. There was an obvious discrepancy between the IGF-I mRNA content within the osteoblast and the free IGF-I level around the osteoblast in either mode of PTH action. The IGF-I might be important for osteoblast-osteoblast communication and bone-forming activity, not only in intermittent PTH administration, but also in the physiological functioning of osteoblasts.