The complete plastome genome sequence of Cynanchum otophyllum (Asclepiadaceae), a unique medicinal species in China.

Cynanchum otophyllum Schneid is an important medicinal plant in China. In this paper, the chloroplast genome of C. otophyllum was sequenced based on high-throughput technology, and the chloroplast genome structure characteristics and phylogenetic relationship of C. otophyllum were analyzed. The results showed the complete plastome genome size of C. otophyllumis 160,874bp, including one small single copy (SSC, 19,851bp) and one large single copy (LSC, 92,009bp) regions isolated by a pair of inverted repeat regions (IRs, 24,507bp). The whole plastome genome including 84 protein encoding genes, 8 rRNA and 37 tRNA. Based on the phylogenetic topologies, C. otophyllum shows close association with additional Gomphocarpus and Asclepias genus. This study contributes to an enhanced understanding of the genetic information of C. otophyllum and provides a theoretical basis for the development of molecular markers and phylogeographic of the species, as well as for constructing the phylogenetic tree of Asclepiadaceae.

Lycium barbarum polysaccharide antagonizes cardiomyocyte apoptosis by inhibiting the upregulation of GRK2 induced by I/R injury, and salvage mitochondrial fission/fusion imbalance and AKT/eNOS signaling.

Ischemia-reperfusion (I/R) injury is the main reason why infarct size continues to progress during the process of restoring myocardial perfusion, and it significantly increases the risk of death. At present, the therapeutic effects of clinically used drugs are limited. Therefore, it is particularly necessary to explore myocardial-protective agents that effectively prevent I/R injury. Lycium barbarum polysaccharide (LBP) is a water-soluble polysaccharide extracted from wolfberry fruit. In this study, we found that LBP limited myocardial infarct size, improved adverse remodeling, and reduced cell death and oxidative stress. G protein-coupled receptor kinase-2 (GRK2) is a key molecule involved in myocardial I/R injury. In vivo and in vitro experiments showed that LBP inhibited the upregulation of GRK2 expression induced by I/R injury, which was related to the antiapoptotic effect of LBP. In addition, we found that LBP partially restored I/R-induced mitochondrial fission/fusion imbalance, as well as levels of phosphorylated protein kinase B (p-AKT) and phosphorylated endothelial cell nitric oxide synthase (p-eNOS), and this restorative effect could be attenuated by overexpression of GRK2. Overall, our findings suggest that LBP antagonizes cardiomyocyte apoptosis by inhibiting the upregulation of GRK2 induced by I/R injury and saves mitochondrial fission/fusion imbalance and AKT/eNOS signaling. This study may provide new ideas for the study of I/R injury and the rational application of the herbal medicine LBP.

Selenium attenuates ischemia/reperfusion injury­induced damage to the blood­brain barrier in hyperglycemia through PI3K/AKT/mTOR pathway­mediated autophagy inhibition.

Ischemic stroke is a leading cause of mortality and disability. Diabetes mellitus, characterized by hyperglycemia, is a common concomitant disease of ischemic stroke, which is associated with autophagy dysfunction and blood­brain barrier (BBB) damage following cerebral ischemia/reperfusion (I/R) injury. At present, there is no effective treatment strategy for the disease. The purpose of the present study was to explore the molecular mechanisms underlying the protective effects of selenium on the BBB following I/R injury in hyperglycemic rats. Middle cerebral artery occlusion was performed in diabetic Sprague­Dawley rats. Treatment with selenium and the autophagy inhibitor 3­methyladenine significantly reduced cerebral infarct volume, brain water content and Evans blue leakage, while increasing the expression of tight junction (TJ) proteins and decreasing that of autophagy­related proteins (P<0.05). In addition, selenium increased the phosphorylation levels of PI3K, AKT and mTOR (P<0.05). A mouse bEnd.3 brain microvascular endothelial cell line was co­cultured in vitro with an MA­h mouse astrocyte­hippocampal cell line to simulate the BBB. The cells were then subjected to hyperglycemia, followed by oxygen­glucose deprivation for 1 h and reoxygenation for 24 h. It was revealed that selenium increased TJ protein levels, reduced BBB permeability, decreased autophagy levels and enhanced the expression of phosphorylated (p)­AKT/AKT and p­mTOR/mTOR proteins (P<0.05). Treatment with wortmannin (an inhibitor of PI3K) significantly prevented the beneficial effects of selenium on the BBB, whereas insulin­like growth factor 1 (a PI3K activator) mimicked the effects of selenium. In conclusion, the present findings indicated that selenium can inhibit autophagy by regulating the PI3K/AKT/mTOR signaling pathway, significantly preventing BBB damage following cerebral I/R injury in hyperglycemic conditions.

Characterization of the complete chloroplast genome of Aconitum pendulum (Ranunculaceae), an endemic medicinal herb.

Aconitum pendulum is an endemic medicinal herb, which has effects of relieving pains and is widely used to treat frostbite, rheumatoid arthritis, and so on. In this study, we sequenced the complete chloroplast genome with Illumina sequencing technology. The complete chloroplast genome length is 155,597 bp, shows a typical tetrad structure, which manifests as one large and one small single copy (LSC and SSC) regions of 86,336 and 16,961 bp, isolated by two inverted repeat regions (IRs) of 26,150 bp. This study annotated altogether 131 unique genes, consisting of 86 protein-encoding genes, 8 rRNA, and 37 tRNA. According to the maximum likelihood phylogenetic tree based on 20 complete chloroplast genomes, A. pendulum shows close association with additional Aconitum subgenus. The chloroplast genome-wide for A. pendulum would help to conserving the precious natural populations.

The complete chloroplast genome of Polygonatum odoratum (Liliaceae), an endemic medicinal herb.

In this study, we sequenced the complete chloroplast genome of Polygonatum odoratum with Illumina sequencing technology. The complete chloroplast genome length is 156,082bp, shows a typical tetrad structure, which manifests as one large and one small single copy (LSC and SSC) regions of 85,009 and 18,513bp, isolated by two inverted repeat regions (IRs) of 26,280bp. This study annotated altogether 131 unique genes, consisting of 86 protein-encoding genes, 8 rRNA and 38tRNA. According to the maximum likelihood phylogenetic tree based on 8 complete chloroplast genomes, P. odoratum shows close association with additional Maianthemum genus. The chloroplast genome-wide for P. odoratum would help to conserving the precious natural populations.

The complete chloroplast genome of Polygonatum odoratum (Liliaceae), an endemic medicinal herb.

In this study, we sequenced the complete chloroplast genome of Polygonatum odoratum with Illumina sequencing technology. The complete chloroplast genome length is 156,082 bp shows a typical tetrad structure, which manifests as one large and one small single copy (LSC and SSC) regions of 85,009 and 18,513 bp, isolated by two inverted repeat regions (IRs) of 26,280 bp. This study annotated altogether 131 unique genes, consisting of 86 protein-encoding genes, 8 rRNA, and 38 tRNA. According to the maximum likelihood phylogenetic tree based on eight complete chloroplast genomes, P. odoratum shows a close association with additional Maianthemum genus. The chloroplast genome-wide for P. odoratum would help to conserve the precious natural populations.

FoxA2 inhibits the proliferation of hepatic progenitor cells by reducing PI3K/Akt/HK2-mediated glycolysis.

FoxA2 is an essential transcription factor for liver organogenesis and homeostasis. Although reduced expression of FoxA2 has been associated with chronic liver diseases, hepatic progenitor cells (HPCs) that are activated in these circumstances express FoxA2. However, the functional effects and underlying mechanism of FoxA2 in HPCs are still unknown. As revealed by immunostaining, HPCs expressed FoxA2 in human cirrhotic livers and in the livers of choline-deficient diet supplemented with ethionine (CDE) rats. Knocking down FoxA2 in HPCs isolated from CDE rats significantly increased cell proliferation and aerobic glycolysis. Moreover, gene transcription, protein expression, and the enzyme activities of hexokinase 2 (HK2) were upregulated, and blocking HK2 activities via 2-deoxyglucose markedly reduced cell proliferation and aerobic glycolysis. Kyoto Encyclopedia of Genes and Genomes analysis revealed that FoxA2 knockdown enhanced the transcription of genes involved in the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway and triggered downstream Akt phosphorylation. Blocking the PI3K/Akt pathway by Ly294002 inhibited HK2 activities, aerobic glycolysis, and cell proliferation in FoxA2-knockdown cells. Therefore, FoxA2 plays an important role in the proliferation and inhibition of HPCs by suppressing PI3K/Akt/HK2-regulated aerobic glycolysis.

Acquisition of exogenous fatty acids renders apicoplast-based biosynthesis dispensable in tachyzoites of Toxoplasma.

Toxoplasma gondii is a common protozoan parasite that infects a wide range of hosts, including livestock and humans. Previous studies have suggested that the type 2 fatty acid synthesis (FAS2) pathway, located in the apicoplast (a nonphotosynthetic plastid relict), is crucial for the parasite's survival. Here we examined the physiological relevance of fatty acid synthesis in T. gondii by focusing on the pyruvate dehydrogenase complex and malonyl-CoA-[acyl carrier protein] transacylase (FabD), which are located in the apicoplast to drive de novo fatty acid biosynthesis. Our results disclosed unexpected metabolic resilience of T. gondii tachyzoites, revealing that they can tolerate CRISPR/Cas9-assisted genetic deletions of three pyruvate dehydrogenase subunits or FabD. All mutants were fully viable in prolonged cultures, albeit with impaired growth and concurrent loss of the apicoplast. Even more surprisingly, these mutants displayed normal virulence in mice, suggesting an expendable role of the FAS2 pathway in vivo Metabolic labeling of the Δpdh-e1α mutant showed reduced incorporation of glucose-derived carbon into fatty acids with medium chain lengths (C14:0 and C16:0), revealing that FAS2 activity was indeed compromised. Moreover, supplementation of exogenous C14:0 or C16:0 significantly reversed the growth defect in the Δpdh-e1α mutant, indicating salvage of these fatty acids. Together, these results demonstrate that the FAS2 pathway is dispensable during the lytic cycle of Toxoplasma because of its remarkable flexibility in acquiring fatty acids. Our findings question the long-held assumption that targeting this pathway has significant therapeutic potential for managing Toxoplasma infections.

[Expression of Cav-1, AQP1 and AQP5 in lung of acute pancreatitis-associated lung injury rats and the therapeutic role of Qingyitang].

OBJECTIVE: to investigate the expression and location of caveolin-1 (Cav-1), aquaporin 1 (AQP1) and AQP5 in the lung of acute pancreatitis-associated lung injury rats and to determine the role of these molecules in the pathologic progress and the potential therapeutic mechanisms of Qingyitang. METHODS: forty Wistar rats were randomly divided into sham operation (SHAM) group, acute lung injury (ALI) group, dexamethasone (DEX) group and Qingyitang (QYT) group. ALI was induced by reverse injection of deoxycholate into biliopancreatic duct of rats. Blood and lung tissues were collected after 24 h. Serum amylase, lung wet/dry (W/D) ratio and pathological section were detected to evaluate the degree of lung injury. reverse transcription-polymerase chain reaction was taken to detect the mRNA levels of Cav-1, AQP1 and AQP5. Lipid rafts were prepared for detection of the distribution and expression level of Cav-1, AQP1 and AQP5 proteins by Western blot. RESULTS: the concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously increased in ALI rats. Cav-1, AQP1 and AQP5 were present in the lung while the mRNA level decreased in ALI rats. Cav-1 appeared mainly in lipid rafts and less in non-lipid rafts. AQP1 was localized to lipid rafts while AQP5 to non-lipid rafts. The localization of these three molecules in the lung of ALI rats did not change compared with SHAM rats while their protein levels decreased. Compared with ALI rats, the concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously decreased in DEX and QYT rats. The mRNA and the protein expression of Cav-1, AQP1 and AQP5 increased in various degrees by DEX or QYT treatment. CONCLUSION: Cav-1 and AQP1 are enriched in lipid rafts while AQP5 in non-lipid rafts. The down-regulated expression of these three molecules may play important role in acute pancreatitis-associated lung injury. DEX and QYT may relieve lung injury effectively by up-regulating the expressions of Cav-1, AQP1 and AQP5.

[Study on rapid reproduction of Scutellaria amoena].

Tissue cultures of Scutellaria amoena were established from the young stem explants on MS + IAA 0.1 mg/L + BA 0. 5 (0.2) mg/L. Propagation cultures were established on MS + BA 0.2 (1.5) mg/L + IAA 0.1 mg/L + NAA 0.1 mg/L + 2, 4-D 0.5 (1.5) mg/L. Rooting of planlet was established on 1/2 MS + NAA 0.1 (0.2) mg/L + IBA 0.1 (0.2) mg/L + PP333 1.0 mg/L and the rate can reach 100%.

Inhibitory effects of anti-SARS traditional Chinese medicines on the UV irradiation of lambda-lysogen.

By using lambda-lysogen as a model, the inhibitory effects of anti-severe acute respiratory syndrome (SARS) traditional Chinese medicines (TCMs) prescription I on the UV irradiation were investigated in this present study. It was found that the prescription I possessed obvious inhibitory effects on the UV induction of lambda-lysogen, the inhibitory rate reaching 83.87%. Among five medicinal herbs prescribed in that formula, Herba Patriniae, Radix Astragali and Radix Glycyrrhizae played important roles. When these three herbs were eliminated from the recipe separately, the inhibitory effects were prominently decreased. If only one of these five medicinal herbs was added into the medium of lambda-lysogen, the inhibitory rates ranged from 27.0% approximately 45.0%. By electron spin resonance (ESR) detection, we found that the prescription I, Herba Patriniae and other main herbs in that recipe, could quench effectively the free radicals generated in the process of lambda-lysogenic cells by UV. These results provide a novel idea for further studying the pharmacology of TCM and exploring the mechanism of SARS virus infection.