Identification and expression analysis of expansin gene family in Salvia miltiorrhiza.

BACKGROUND: Expansins (EXP) are important enzymes that are involved in the extension of plant cells and regulation of root configurations, which play important roles in resisting various stresses. As a model medicinal plant, Salvia miltiorrhiza is well recognized for treating coronary heart disease, myocardial infection, and other cardiovascular and cerebrovascular diseases; however, the SmEXP gene family has not yet been analyzed. METHODS: The SmEXP family was systematically analyzed using bioinformatics. Quantitative real-time PCR was employed to analyze the tissue expression patterns of the SmEXP family, as well as its expression under abscisic acid (ABA) treatment and abiotic stress. Subcellular localization assay revealed the localization of SmEXLA1, SmEXLB1, and SmEXPA2. RESULTS: This study identified 29 SmEXP that belonged to four different subfamilies. SmEXP promoter analysis suggested that it may be involved in the growth, development, and stress adaptation of S. miltiorrhiza. An analysis of the expression patterns of SmEXP revealed that ABA, Cu2+, and NaCl had regulatory effects on its expression. A subcellular localization assay showed that SmEXLA1 and SmEXLB1 were located on the nucleus and cell membrane, while SmEXPA2 was located on the cell wall. CONCLUSION: For this study, the SmEXP family was systematically analyzed for the first time, which lays a foundation for further elucidating its physiological and biological functionality.

Electroacupuncture Ameliorates Depression-Like Behaviors Comorbid to Chronic Neuropathic Pain via Tet1-Mediated Restoration of Adult Neurogenesis.

Although electroacupuncture (EA) stimulation is a widely used therapy for chronic pain and comorbid psychiatric disorders, its long-term effects on chronic neuropathic pain-induced depression and the underlying mechanisms remain elusive. In the present study, we found that EA stimulation was able to restore adult neurogenesis in the ventral dentate gyrus (DG), by both increasing neuronal differentiation and restoring the normal morphology of newborn dendrites, in mice with spared nerve injury surgery. By ablating the Nestin+ neural stem cells (NSCs) via diphtheria toxin fragment A expression, we further proved that neurogenesis in the ventral DG was crucial to the long-term, but not the immediate antidepressant effect of EA, nor was it associated with nociception. Furthermore, we found that the restoration of neurogenesis was dependent on Tet1-mediated epigenetic modification upon EA treatment. Tet1 could bind to the promoter of the Prox1 gene, thus catalyzing its demethylation and facilitating its expression, which finally contributed to the restoration of neurogenesis and amelioration of depression-like behaviors induced by chronic neuropathic pain. Thus, we conclude that EA stimulation restores inhibited Tet1 expression in hippocampal NSCs of mice with chronic neuropathic pain, and increased Tet1 expression ameliorates hypermethylation of Prox1 and restores normal adult neurogenesis in the ventral DG, which contributes to the long-term antidepressant effect of EA.

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.

SmMYB4 Is a R2R3-MYB Transcriptional Repressor Regulating the Biosynthesis of Phenolic Acids and Tanshinones in Salvia miltiorrhiza.

Salvia miltiorrhiza Bunge is one of the most famous traditional Chinese medicinal plants. The two most important classes of pharmaceutically relevant compounds in S. miltiorrhiza are phenolic acids and tanshinones. The MYB family of transcription factors may efficiently regulate the secondary metabolism in plants. In this study, a subgroup 4 R2R3MYB transcription factor gene, SmMYB4, was isolated from S. miltiorrhiza and functionally characterized using overexpression and a RNAi-mediated silencing. We achieved a total of six overexpressions and eight RNAi transgenic lines from the Agrobacterium leaf disc method. The content of the total phenolics, rosmarinic acid, and salvianolic acid B markedly decreased in the SmMYB4-overexpressing lines but increased in the SmMYB4-RNAi lines. The content of the total tanshinones, cryptotanshinone, and tanshinone IIA decreased in the SmMYB4-overexpressing transgenic lines but increased in the SmMYB4-RNAi lines. A gene expression analysis demonstrated that SmMYB4 negatively regulated the transcription of the critical enzyme genes involved in the phenolic acid and tanshinone biosynthesis. The genetic control of this transcriptional repressor may be used to improve the content of these bioactive compounds in the cultivated S. miltiorrhiza.

Leonurine: From Gynecologic Medicine to Pleiotropic Agent.

Leonurus japonicus Houtt, a Chinese traditional herbal medicine, has been widely used to cure gynecological diseases, such as incomplete abortion and menoxenia. Leonurine, a major active alkaloid compound only be found in Leonurus japonicus Houtt, has been successfully extracted and purified. Recent evidence has shown that leonurine can regulate a variety of pathologic processes including oxidative stress, inflammation, fibrosis, apoptosis, and multiple metabolic disorders. Here, we have reviewed the pharmacological actions and biological functions of leonurine, with a focus on the role of leonurine in the amelioration of various pathological processes. Insights into the related signaling pathways and molecular mechanisms have strengthened our understanding on the function of leonurine in the alleviation of multiple pathological states. Our summary of the existing researches should help direct future research into the basic science and clinical applications in related diseases.

Selenium decreases methylmercury and increases nutritional elements in rice growing in mercury-contaminated farmland.

Methylmercury (MeHg) in rice grains grown in Hg-contaminated areas has raised environmental health concerns. Pot experiments found that selenium (Se) could reduce MeHg levels in rice grains. However, relatively high levels of Se (up to 6 mg/kg) were applied in these pot experiments, which may have adverse effects on the soil ecology due to the toxicity of Se. The aims of this work were thus to study 1) the effect of low levels of Se on the accumulation and distribution of Hg, especially MeHg, in rice plants grown in a real Hg-contaminated paddy field and 2) the effect of Se treatment on Se and other nutritional elements (e.g., Cu, Fe, Zn) in grains. A field study amended with different levels of Se was carried out in Hg-contaminated paddy soil in Qingzhen, Guizhou, China. The levels of MeHg and total Hg were studied using cold vapor atomic fluorescence spectrometry (CVAFS) and inductively coupled plasma mass spectrometry (ICP-MS). The distribution and relative quantification of elements in grains were examined by synchrotron radiation X-ray fluorescence analysis (SR-XRF). This field study showed that low levels of Se (0.5 µg/mL, corresponding to 0.15 mg Se/kg soils) could significantly reduce total Hg and MeHg in rice tissues. Se treatment also reduced Hg distribution in the embryo and endosperm and increased the levels of Fe, Cu, Zn and Se in grains and especially embryos. This field study implied that treatment with an appropriate level of Se is an effective approach to not only decrease the level of MeHg but to also increase the levels of nutritional elements such as Fe, Cu, Zn and Se in rice grains, which could bring beneficial effects for rice-dependent residents living in Hg-contaminated areas.

Botanic Metallomics of Mercury and Selenium: Current Understanding of Mercury-Selenium Antagonism in Plant with the Traditional and Advanced Technology.

The antagonistic effect between mercury (Hg) and selenium (Se) is conclusively established in animals and human beings in the past decades. However, the underlying mechanisms of the interactions between Hg and Se in plants, as well as the metabolism of Hg-Se compounds in crops are still far from being understood. The botanic metallomics of Hg and Se mainly focuses on the translocation, transformation, and metabolism of Hg and Se in the environmental and botanic systems employing metallomics methods. An adequate understanding of the biological behavior of Hg and Se in plant is beneficial for sequestration of Hg and Se in soil-plant systems with high Hg and Se contamination. It can also provide a molecular mechanistic basis for Se supplementation in Se-deficient areas. Here, the key developments in current understanding of Hg and Se interactions in plants are reviewed. The metabolism and antagonism of Hg and Se in various plants, as well as the advanced analytical methods commonly used in this field, are summarized and discussed. As suggested, plant Hg and Se uptake, metabolism, and antagonism can be taken into account for detoxification and remediation strategies for the reduction of Hg and Se in the food chain.

Nanoelemental selenium alleviated the mercury load and promoted the formation of high-molecular-weight mercury- and selenium-containing proteins in serum samples from methylmercury-poisoned rats.

Selenite (Se4+) has been found to counteract the neurotoxicity of methylmercury (MeHg) in MeHg-poisoned rats. However, Se4+ has narrow range between its toxic and beneficial effects. Nanoelemental selenium (SeNPs) was found to be less toxic than other forms of Se such as Se4+. In this study, the effects of SeNPs on the load of mercury (Hg) in rats were investigated. Hyphenated technique based on size-exclusion chromatography coupled with UV and inductively coupled plasma mass spectrometry (SEC-ICP-MS) detection and synchrotron radiation X-ray fluorescence spectroscopy (SR-XRF) were used to analyze the Hg-Se-containing proteins in the serum from MeHg-poisoned rats. The Hg-Se-containing fractions monitored by UV and ICP-MS were further characterized by MALDI-TOF-MS. Elevated serum Hg and Se levels were found in MeHg-poisoned rats after SeNPs treatment. Three main Hg-containing bands with molecular weights (MWs) of 25, 62 and 140 kDa were detected in the control samples. Treatment with SeNPs increased the Hg content in proteins at 62 and 170 kDa and decreased the Hg content at 25 kDa. The fraction with 25 kDa was assigned to metallothioneins (MTs), and fractions with 40 and 75 kDa were assigned to albumin. This study showed that the low-toxicity SeNPs could reduce the Hg load in the tissues and promote the formation of high molecular weight Hg- and Se-containing proteins in MeHg-poisoned rats.

The effect of dietary supplementation of low crude protein on intestinal morphology in pigs.

To explore the effects of reducing the Cp levels on intestinal barrier function, low Cp (LP) and NRC standard Cp (NP) diets were fed to pigs from 45 to 160 days, and in vitro experiments were performed using monolayers of IPEC-J2 cells. The number of goblet cells, expression of proteins related to cell junction, amino acid transport, glucose transport, transepithelial electrical resistance (TEER), dextran permeability, and IL-6 secretion level were detected in pigs. The results demonstrated that a moderate reduction of Cp levels did not affect intestinal morphology, as demonstrated by a normal villi height, crypt depth and normal numbers of goblet cells. The maintenance of the intestinal structure obtained with LP was also confirmed by stable mRNA expression levels of muc2 and E-cadherin in the jejunum. We also found that LP did not affect the protein expression of cationic amino acid transporter 1 (CAT-1) and alanine serine cysteine transporter 1 (ASCT1) from 45 to 160 days. Moreover, the excitatory amino acid transporter 3 (EAAT3), sodium-glucose cotransporter 1 (SGLT1) and glucose transporter (GLUT2) protein expression levels in the jejunum were significantly increased at a certain age during the rearing period. Furthermore, we also demonstrated that a reduction in protein concentration up to 15% in the cultural medium of IPEC-J2 cells did not impact the mucosal barrier function. This study demonstrated that a moderate reduction of the protein level did not affect intestinal mucosal barrier function and morphology in the jejunum.

Ethanol Extract from Brucea Javanica Seed Inhibits Angiogenesis Mediated by Platelet Derived Growth Factor Receptor-beta.

The present study aimed to investigate the effects of ethanol extract from Brucea javanicaseed (EEBJS) on the angiogenesis of human umbilical vein endothelial cells (HUVECs) and the possible molecular signal involved. Firstly, a Matrigel-based in vitro angiogenesis assay demonstrated that EEBJS inhibited the angiogenesis of HUVECs in a dose-dependent manner. Then by using porcine aortic endothelial cells which stably express human PDGFR-beta, we found that the inhibition of angiogenesis was mediated by PDGFR-beta. Taken together, we conclude that EEBJS inhibited the angiogenesis function of the vascular endothelial cells mediated by PDGFR-beta, and postulate that it might contribute to the therapeutic effects of EEBJS on malignant tumors.

The Effect of Mindfulness Training on Proactive and Reactive Cognitive Control.

Previous studies have demonstrated that mindfulness practice can improve general cognitive control. However, little research has examined whether mindfulness practices affect different cognitive control strategies. According to the dual mechanisms of control (DMC) model, different cognitive control strategies may play distinct roles in individuals' lives. Proactive control allows people to maintain and prepare for goals, whereas reactive control allows them to respond flexibly to a changing environment. Thus, this study investigates the influences of mindfulness training on proactive and reactive control measured by the AX version of the Continuous Performance Test (AX-CPT). Thirty participants completed AX-CPT and the Five Facet Mindfulness Questionnaire (FFMQ) before and after random assignment to either an 8-week mindfulness training group or a control group. The results showed no interaction between group and test time for AY or BX trial type, but the training group had fewer post-test errors on the BX trial and a higher Behavior Shift Index (BSI) of reaction time (RT) compared with the control group. This finding indicates enhanced trend of proactive control with mindfulness training. A positive correlation between the BSI of RT and observing scores on the FFMQ confirmed the connection between attentional components in mindfulness and proactive control. Errors on the AY trial in the post-test decreased in both groups, reflecting reactive control that did not differ between groups. The 8-week mindfulness training demonstrates a potential improvement effect on proactive control and could be helpful in overcoming interference.

Evidence for molecular antagonistic mechanism between mercury and selenium in rice (Oryza sativa L.): A combined study using 1, 2-dimensional electrophoresis and SR-XRF techniques.

Mercury (Hg) is a hazardous chemical in the environment and can accumulate in the food chain. Selenium (Se) is a necessary element for human health and has antagonistic effects on Hg toxicity. In this work, we investigated the effect of Se on Hg containing and Hg-responsive proteins in rice using 1, 2-dimensional electrophoresis combined with SR-XRF techniques. Two weeks old rice seedlings were exposed to Hg and/or Se compounds. After 21days proteins in the rice roots were separated by electrophoresis and their metal contents were determined by X-ray fluorescence to identify Hg and Se responsive biomolecules. The results show that under Hg stress alone Hg is bound to proteins with molecular weights of 15-25kDa. With the addition of Se, a new Hg-containing protein band in the 55-70kDa range was also found, while the content of Hg in the 15-25kDa proteins decreased. Ten and nine new protein spots were identified after adding Se to inorganic Hg and methylmercury exposed roots, respectively. Adding Se regulates the abundance of proteins associated with carbohydrate and energy metabolism, stress response, cell cycle, and DNA replication indicating that these proteins mediate the antagonism of Se against Hg toxicity. This study helps us to better understand the molecular mechanism of Hg tolerance as well as the molecular antagonism between Hg and Se in rice plants.

Elevated mercury bound to serum proteins in methylmercury poisoned rats after selenium treatment.

Methylmercury is a toxic pollutant and is generated by microbial methylation of elemental or inorganic mercury in the environment. Previous study found decreased hepatic MDA levels and urinary mercury levels in methylmercury poisoned rats after sodium selenite treatment. This study further found increased mercury levels in serum samples from methylmercury poisoned rats after selenium treatment. By using size exclusion chromatography coupled to inductively coupled plasma mass spectrometry, three Hg- binding protein fractions and two Se-binding protein fractions were identified with the molecular weight of approximately 21, 40, and 75 kDa and of 40 and 75 kDa, respectively. Elevated mercury level in the 75 kDa protein fraction was found binding with both Hg and Se, which may explain the decreased urinary Hg excretion in MeHg poisoned rats after Se treatment. MALDI-TOF-MS analysis of the serum found that the 75 kDa protein fractions were albumin binding with both Hg and Se and the 21 kDa fraction was Hg- binding metallothionein.

Selenium modulates mercury uptake and distribution in rice (Oryza sativa L.), in correlation with mercury species and exposure level.

Rice cultured in Hg- and/or Se-contaminated fields is an important food source of human Hg/Se intake. There are elevated Hg and Se levels in the soil of the Wanshan District, Guizhou Province. Here we attempted to explore how a Hg antagonist, Se, modulates the absorption and accumulation of inorganic mercury (IHg) and methylmercury (MeHg) in rice. The effects of Se on the content and transportation of Hg in hydroponic and soil cultured rice plants were examined. The results show that IHg mainly accumulated in the rice roots, but some also accumulated in the rice grain. In comparison to IHg, MeHg can be concentrated in the rice grain, and the proportion of MeHg in the rice grain may account for above 40% of the total Hg. Se can protect against Hg phytotoxicity in rice and inhibit IHg accumulation in rice tissues, but was not remarkable for MeHg at a low dosage exposure level in this study. These discrepancies imply mechanistic differences between IHg and MeHg absorption and accumulation in rice. This study illustrates that Se plays an important role in modulating Hg uptake, transportation and accumulation in rice. Therefore, Se is considered to be a naturally existing element that effectively reduces Hg accumulation in rice, which may have significant implications for food safety.

Efficacy and safety of berberine for congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy.

This study was designed to assess the efficacy and safety of berberine for chronic congestive heart failure (CHF). One hundred fifty-six patients with CHF and >90 ventricular premature complexes (VPCs) and/or nonsustained ventricular tachycardia (VT) on 24-hour Holter monitoring were randomly divided into 2 groups. All patients were given conventional therapy for CHF, consisting of angiotensin-converting enzyme inhibitors, digoxin, diuretics, and nitrates. Patients in the treatment group (n = 79) were also given berberine 1.2 to 2.0 g/day. The remaining 77 patients were given placebo. Symptoms, a 6-minute walk test, left ventricular (LV) ejection fraction (EF), frequency and complexity of VPCs, and quality of life were assessed after 8 weeks of treatment and during a mean 24-month follow-up. After treatment with berberine, there was a significantly greater increase in LVEF, exercise capacity, improvement of the dyspnea-fatigue index, and a decrease of frequency and complexity of VPCs compared with the control group. There was a significant decrease in mortality in the berberine-treated patients during long-term follow-up (7 patients receiving treatment died vs 13 on placebo, p <0.02). Proarrhythmia was not observed, and there were no apparent side effects. Thus, berberine improved quality of life and decreased VPCs and mortality in patients with CHF.