Search

Ethnopharmacological relevance Hanchuan Zupa Granule (HCZP), a traditional Chinese ethnodrug, has the functions of supressing a cough, resolving phlegm, warming the lungs, and relieving asthma. In clinical practice employing traditional Chinese medicine (TCM), HCZP is commonly used to treat acute colds, cough and abnormal mucous asthma caused by a cold, or “Nai-Zi-Lai” in the Uygur language. Studies have confirmed the use of HCZP to treat cough variant asthma (CVA) and other respiratory diseases. However, the pharmacological mechanisms of HCZP remain unrevealed. Aim of the study To investigate the anti-tussive and anti-asthmatic effects and the possible pharmacological mechanisms of HCZP in the treatment of CVA. Materials and methods A guinea pig CVA animal model was established by intraperitoneal injection of ovalbumin (OVA) combined with intraperitoneal injection of aluminium hydroxide adjuvant and atomized OVA. Meanwhile, guinea pigs with CVA received oral HCZP (at dosages of 0.571, 0.285 and 0.143 g/kg bodyweight). The number of coughs induced by aerosol capsaicin was recorded, and the airway hyperresponsiveness (AHR) of CVA guinea pigs was detected with the FinePointe series RC system. H&E staining of lung tissues was performed to observe pathological changes. ELISA was used to detect inflammatory cytokines. qRT-PCR and western blotting analyses were used to detect the expression of Th1-specific transcription factor (T-bet), Th2-specific transcription factor (GATA3), and Toll-like receptor 4 (TLR4) signal transduction elements. These methods were performed to assess the protective effects and the potential mechanisms of HCZP on CVA. Results Great changes were found in the CVA guinea pig model after HCZP treatment. The number of coughs induced by capsaicin in guinea pigs decreased, the body weights of guinea pigs increased, and inflammation of the eosinophilic airway and AHR were reduced simultaneously. These results indicate that HCZP has a significant protective effect on CVA. A pharmacological study of HCZP showed that the levels of interleukin-4 (IL-4) and IL-5 and tumour necrosis factor-α (TNF-α) in serum decreased. The amount of interferon-γ (IFN-γ) increased, mRNA and protein expression of TLR4 and GATA3 weakened, and mRNA and protein expression of T-bet increased. Conclusions HCZP ameliorated the symptoms of guinea pigs with CVA induced by OVA by regulating the Th1/Th2 imbalance and TLR4 receptors.

Ethnopharmacological relevance Zhizhu Xiang (ZZX for short) is the root and rhizome of Valeriana jatamansi Jones, which is a Traditional Chinese Medicine (TCM) used to treat various mood disorders for more than 2000 years, especially anxiety. However, there have been few investigations to clarify the compounds in ZZX for the treatment of anxiety. Aim of the study Our previous study has identified five anti-anxiety components, including hesperidin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C and chlorogenic acid, from extract of ZZX. In order to find the optimal combination and the underlying mechanism of these five components in the treatment of anxiety disorder, researches were designed based on uniform design method and proteomic technology. Materials and methods The samples with different proportion and content of the five active components were arranged by uniform design method. Then a mathematical model was formulated using partial least square method and stepwise regression analysis. Moreover, the empty bottle stress-induced anxiety rat model was established, and the anti-anxiety effect was recorded by the unconditioned reflex elevated maze test and the open field test. In addition, the isobaric tags for relative and absolute quantitation (iTRAQ) technique, along with the multidimensional liquid chromatography and high-resolution mass spectrometry were applied in proteomic study. At last, the result of proteomic analysis was further confirmed by Western blot. Results The optimal combination of the components from the extract of ZZX was 1.153 mg/kg hesperidin, 2.197 mg/kg Isochlorogenic acid A, 0.699 mg/kg Isochlorogenic acid B and 1.249 mg/kg Chlorogenic acid. Total 6818 proteins were identified using proteomic analysis and 80 differentially expressed proteins were used for further bioinformatic analysis. These proteins were involved in the neuroactive ligand-receptor interaction, protein digestion and absorption, cholesterol metabolism, Chagas disease, and AGE/RAGE signaling pathway. Conclusions The composition and proportion of anti-anxiety components in extract of ZZX was disclosed, and there was an anti-anxiety effect for the combined components of flavonoids and phenolic acids. Through proteomic analysis and Western blot, it was found that the effective components of extract of ZZX can exert synergistic anti-anxiety effects via the regulation of multi-signaling pathways. These findings could provide a preliminary research basis for the development of new low-toxic, efficient, stable and controllable anti-anxiety drugs.

Larix kaempferi is widely cultivated because of its significant ecological and economic values. However, the molecular mechanism underlying the growth and development of Larix kaempferi remains largely unknown because of its long growth cycle and highly complex genetic background. In this study, global transcriptome sequencing of Larix kaempferi was conducted via Solexa sequencing. In total, 15.49 million clean reads were assembled into 20,395 unigenes and then annotated. The study found hundreds of highly expressed unigenes involved in growth and development (e.g. plant hormone regulation pathways or starch and sucrose metabolic pathways), of which 25 unigenes with full-length coding regions interested us the most. In addition, two gene families containing the zinc finger and nucleotide-binding site (NBS) domains were further explored. A total of 48 C3H-type zinc finger genes, most containing a conversed C-X7–10-C-X5-C-X3-H motif, were identified and divided into eight distinct subfamilies through phylogenetic analysis. Twenty-one genes containing the NBS domain represented four subfamilies. These findings provided new insights into the transcriptome characteristics of Larix kaempferi and abundant sequence information to elucidate the roles of the functional genes in Larix kaempferi.

Drought decreases crop productivity more than any other type of environmental stress. Transcription factors (TFs) play crucial roles in regulating plant abiotic stress responses. The Arabidopsis thaliana gene DREB1A/CBF3, encoding a stress-inducible TF, was introduced into Salvia miltiorrhiza Ectopic expression of AtDREB1A resulted in increased drought tolerance, and transgenic lines had higher relative water content and Chl content, and exhibited an increased photosynthetic rate when subjected to drought stress. AtDREB1A transgenic plants generally displayed lower malondialdehyde (MDA), but higher superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities under drought stress. In particular, plants with ectopic AtDREB1A expression under the control of the stress-induced RD29A promoter exhibited more tolerance to drought compared with p35S::AtDREB1A transgenic plants, without growth inhibition or phenotypic aberrations. Differential gene expression profiling of wild-type and pRD29A::AtDREB1A transgenic plants following drought stress revealed that the expression levels of various genes associated with the stress response, photosynthesis, signaling, carbohydrate metabolism and protein protection were substantially higher in transgenic plants. In addition, the amount of salvianolic acids and tanshinones was significantly elevated in AtDREB1A transgenic S. miltiorrhiza roots, and most of the genes in the related biosynthetic pathways were up-regulated. Together, these results demonstrated that inducing the expression of a TF can effectively regulate multiple genes in the stress response pathways and significantly improve the resistance of plants to abiotic stresses. Our results also suggest that genetic manipulation of a TF can improve production of valuable secondary metabolites by regulating genes in associated pathways.

Tanshinones are compounds of diterpenoid, which are used in the treatment of cardiovascular diseases; however there is an urgent need to improve their production using biotechnological methods. In the present study, Agrobacterium-mediated transformation was used to produce hairy roots of Salvia miltiorrhiza expressing kaurene synthase-like (SmKSL), a tanshinone biosynthesis gene. The results revealed that the yield of total tanshinone increased by a maximum of 2.7-fold in the transgenic Salvia miltiorrhiza hairy roots overexpressing SmKSL as compared with wild-type control lines. Treatment with elicitors further increased the total tanshinone production in the SmKSL-overexpressing lines. Yeast extract treatment of line p35S::SmKSL-15 produced the highest total tanshinone content (4.2 mg/g dry weight), representing an increase of 3.5-fold compared with that in the wild-type control lines. At the same time, qRT-PCR analysis indicated that tanshinone accumulation correlated positively with the transcriptional level of the two pathway genes.

Grain boundary diffusion process (GBDP) using Pr60Tb10Cu30−xAlx (x = 0, 5, 15, 25, 30, atomic fraction) alloy ribbons, named as A0–A30 alloys, was applied to high maximum energy product (BH)max Nd-Fe-B sintered magnet. After diffusion, coercivity (Hci) of A0 magnet is significantly enhanced from the original 11.4 kOe to 19.5 kOe with remanence (Br) decreasing from 14.65 kGs to14.32 kGs, which is mainly attributed to magnetic hardening effect by Tb-rich shells. With substituting 5 at% Al for Cu, the Hci and Br of A5 magnet are simultaneously enhanced by the homogenized distribution of Tb-rich shells and the reduced interface energy of Nd-rich phase and Nd2Fe14B phase. The highest coercivity of 22.4 kOe is obtained for A25 magnet by further strengthening magnetic isolation of Nd2Fe14B phase, however, which results in a drastic decrease in Br due to the increased fraction of Nd-rich phase and too much substitution of Al for Fe in Nd2Fe14B phase. Therefore, the Hci (kOe) + (BH)max (MGOe) reaches to the peak value for A5 magnet. Also, it is proven that Cu/Al co-addition can effectively improve Tb diffusion efficiency by modifying intergranular Nd-rich phase.

The incidence of type 1 diabetes mellitus (T1DM) is increasing year by year, gut microbiota is considered to be closely related to the occurrence and development of T1DM in recent years. In this study, Sprague Dawley (SD) rats were intraperitoneally injected with 75mg/kg streptozotocin to establish T1DM model, fecal samples were collected and DNA were extracted, 16S rRNA microbial gene clone library were constructed, and lastly high-throughput sequencing and bioinformatics analysis were performed. The results showed that the abundances of pathogenic bacteria such as Ruminococcaceae, Shigella, Enterococcus, Streptococcus, Rothia and Alistipes associated with infection and inflammation in T1DM rats were up-regulated, while the abundances of beneficial bacteria such as Lactobacillus, Faecalitalea, Butyricicoccus and Allobaculum were reduced. Among them, Butyricicoccus and Allobaculum protect intestinal barrier function by producing short-chain fatty acids. This study suggests that intestinal inflammation and reduction of short chain fatty acids (SCFAs) caused by the imbalance of gut microbiota are crucial to the pathogenesis of T1DM.

In this study, a non-targeted metabolic profiling method based on ultra-performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) was used to characterize the plasma metabolic profile associated with the protective effects of the Sagittaria sagittifolia polysaccharide (SSP) on isoniazid (INH)&mdash, and rifampicin (RFP)-induced hepatotoxicity in mice. Fourteen potential biomarkers were identified from the plasma of SSP-treated mice. The protective effects of SSP on hepatotoxicity caused by the combination of INH and RFP (INH/RFP) were further elucidated by investigating the related metabolic pathways. INH/RFP was found to disrupt fatty acid metabolism, the tricarboxylic acid cycle, amino acid metabolism, taurine metabolism, and the ornithine cycle. The results of the metabolomics study showed that SSP provided protective effects against INH/RFP-induced liver injury by partially regulating perturbed metabolic pathways.

MiR171 plays pleiotropic roles in the growth and development of several plant species. However, the mechanism underlying the miR171-mediated regulation of organ development in broccoli remains unknown. In this study, bol-miR171b was characterized and found to be differentially expressed in various broccoli organs. The ectopic overexpression of bol-miR171b in Arabidopsis affected the leaf and silique development of transgenic lines. In particular, the chlorophyll content of leaves from overexpressed bol-miR171b transgenic Arabidopsis was higher than that of the vector controls. The fertility and seed yield of Arabidopsis with overexpressed bol-miR171b were markedly lower than those of the vector controls. Similarly, overexpressed bol-miR171b transgenic broccoli exhibited dark green leaves with high chlorophyll content, and nearly all of the flowers were sterile. These results demonstrated that overexpression of bol-miR171b could increase the chlorophyll content of transgenic plants. Degradome sequencing was conducted to identify the targets of bol-miR171b. Two members of the GRAS gene family, BolSCL6 and BolSCL27, were cleaved by bol-miR171b-3p in broccoli. In addition to the genes targeted by bol-miR171b-3p, adenylylsulfate reductase 3 ( APSR3), which played important roles in plant sulfate assimilation and reduction, was speculated to be cleaved by bol-miR171b-5p, suggesting that the star sequence of bol-miR171b may also have functions in broccoli. Comparative transcriptome analysis further revealed that the genes involved in chloroplast development and sulfate homeostasis should participate in the bol-miR171b -mediated regulatory network. Taken together, these findings provided new insights into the function and regulation of bol-miR171b in broccoli and indicated the potential of bol-miR171b as a small RNA molecule that increased leaf chlorophyll in plants by genetic engineering.

Background Defining the postoperative pelvic tilt (PT) individually can help to reconstruct sagittal balance. However, the postoperative actual PT is hardly restored to theoretical value. Some cases with theoretical postoperative PT was overcorrected and still did not have normal horizontal visual field after surgery. The objective of this study is to describe the pelvic tilt change after spinal osteotomy in ankylosing spondylitis (AS) kyphotic deformity and evaluate the effect on clinical outcomes. Methods Twenty-three AS patients including 21 men and two women with thoracolumbar kyphosis, who underwent spinal osteotomy from 2013 to 2015 in our center, were retrospectively reviewed. A series of parameters including sacral slop (SS), pelvic incidence (PI), PT, and sagittal vertical axis (SVA) measured on preoperative and postoperative standing radiographs were analyzed. The theoretical postoperative PT (tPT) was calculated by the formula tPT = 0.37 × PI − 7. The radiographic measurements were compared before surgery, 2 weeks and at least 2 years postoperatively. Clinical outcomes were performed with the Oswestry disability index and Scoliosis Research Society-22 surveys. Results Mean age of the patients (2 women, 21 men) was 39.8 ± 9.1 years. Mean follow-up was 27.4 ± 3.8 months, at least 24 months. After spinal osteotomy, SS and SVA were corrected from 11.9° ± 11.2° and 18.0 ± 7.6 mm preoperatively to 25.8° ± 8.1° and 9.6 ± 6.3 mm postoperatively, respectively (p

With the improvement of living standards and a change in lifestyle, the incidence of type 2 diabetes mellitus (T2DM) is increasing. Its etiology is too complex to be completely understand yet. Metabonomics techniques are used to study the changes of metabolites and metabolic pathways before and after the onset of diabetes and make it more possible to further understand the pathogenesis of T2DM and improve its prediction, early diagnosis, and treatment. In this review, we summarized the metabonomics study of T2DM in recent years and provided a theoretical basis for the study of pathogenesis and the effective prevention and treatment of T2DM.

Background Mitochondrial dysfunction has been implicated in the pathogenesis of ischemic heart disease, exacerbating cardiomyocytes injury in myocardial infarction (MI). Peroxisome proliferator-activated receptor gamma co-activator (PGC-1α) has been recognized as the key regulator of mitochondrial biogenesis and energy metabolism. Yiqihuoxue decoction (YQHX), a Traditional Chinese Medicine (TCM) prescription, can prevent and treat ischemic heart disease. However, the mechanisms of YQHX on PGC-1α expression in the ischemic heart have remained unclear. Methods Myocardial ischemia rat model and ischemia/hypoxia injury model in the cardiomyocytes were used to minic human cardiovascular disease. Rats were randomly assigned into 4 groups: Sham, Model, YQHX (8.2 g/kg) and Trimetazidine (10 mg/kg) group. 28 days after MI, cardiac functions and morphology were detected by echocardiography and HE staining, respectively. In vitro, the effects of YQHX on H9c2 cell viability, LDH and ROS were detected, respectively. PGC-1α relevant proteins were evaluated by Western blotting. Results In vivo, echocardiography and HE staining results showed that YQHX improved cardiac functions and modified pathological changes. YQHX enhanced PGC-1α expression and improved the mitochondrial ultrastructure and functions in rats MI model for 4 weeks. Further, we explored its potential mechanisms in cardiomyocytes. In vitro, YQHX significantly enhanced cell viability and reduced LDH release and ROS production induced by hypoxia in cardiomyocytes. Interestingly, exposure of cardiomyocytes to hypoxic conditions for 12 h induced the downregulation of PGC-1α expression, but the expression levels nearly returned to the normal state after hypoxia for 24 h. YQHX significantly enhanced PGC-1α expression between 12 h and 24 h induced by hypoxia through a mechanism associated with the activation of AMPK phosphorylation in H9c2 cells. In addition, YQHX upregulated the expression of Tfam and NRF-1, while NRF-1 expression was completely blocked by an AMPK inhibitor. YQHX largely restored the mitochondrial morphology and increased mitochondrial membrane potential in hypoxia-induced injury. Furthermore, the UHPLC-LTQ-Orbitrap-MSn analysis found that there were 87 chemical constituents in YQHX. Conclusions These results suggest that the protective effect of YQHX on cardiomyocytes against hypoxia-induced injury may be attributed to activation of PGC-1α and maintenance of mitochondrial functions through a mechanism involving the activation of AMPK phosphorylation.

In our previous study, drought-resistant transgenic plants of Salvia miltiorrhiza were produced via overexpression of the transcription factor AtDREB1A. To unravel the molecular mechanisms underpinning elevated drought tolerance in transgenic plants, in the present study we compared the global transcriptional profiles of wild-type (WT) and AtDREB1A-expressing transgenic plants using RNA-sequencing (RNA-seq). Using cluster analysis, we identified 3904 differentially expressed genes (DEGs). Compared with WT plants, 423 unigenes were up-regulated in pRD29A::AtDREB1A-31 before drought treatment, while 936 were down-regulated and 1580 and 1313 unigenes were up- and down-regulated after six days of drought. COG analysis revealed that the ‘signal transduction mechanisms’ category was highly enriched among these DEGs both before and after drought stress. Based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation, DEGs associated with “ribosome”, “plant hormone signal transduction”, photosynthesis”, “plant-pathogen interaction”, “glycolysis/gluconeogenesis” and “carbon fixation” are hypothesized to perform major functions in drought resistance in AtDREB1A-expressing transgenic plants. Furthermore, the number of DEGs associated with different transcription factors increased significantly after drought stress, especially the AP2/ERF, bZIP and MYB protein families. Taken together, this study substantially expands the transcriptomic information for S. miltiorrhiza and provides valuable clues for elucidating the mechanism of AtDREB1A-mediated drought tolerance in transgenic plants.

Ethnopharmacological relevance Arecae semen has been used as vermifuge and digestant in traditional Chinese medicine (TCM) for more than one thousand years. However, the toxicity effect of areca semen and its underlying mechanism are still unclear. The aim of the study: This study was aimed to investigate the toxicity of arecae semen and to explore its mechanisms by serum metabolomics. Materials and methods The male Wistar rats were divided into the control group and treated group (n = 6 in each group), which were given by gavage with distill water or arecae semen aqueous extract (ASAE) once a day for 30 days, respectively. Serum samples were collected from all the rats after treatment of 7-day, 14-day and 30-day for metabolomics analysis. Moreover, biochemistry analysis and histopathological examination were performed at the end of study. Results The phenomenon of diarrhea, less physical activity, tremors and body curl up were observed in the treated group. Additionally, the body weights of treated rats were significantly decreased compared with control rats from the 8th day after oral administration. Except the level of creatinekinase (CK) in the treated group significantly increased compared with the control group, there were no differences on biochemistry parameters and histopathological test in the two groups. Combined with the methods of principal component analysis (PCA), orthogonal projection to latent structure-discrimination analysis (OPLS-DA) and available databases, the treated and control rats were clearly distinguished from each other and 19 metabolites were identified as the potential biomarkers in the arecae semen treated rats. The identified biomarkers indicated that there were perturbations of the phospholipid metabolism, amino acid metabolism and fat acid metabolism in the treated group. Conclusions This indicated that arecae semen possessed certain cardiotoxicity and inhibited the normal growth in Wistar male rats. In addition, the metabolomics approach is a useful tool to study the toxicity in TCM.

Structure identification of chemical constituents of YQHX formula by UHPLC-LTQ Orbitrap MS. There were 87 peaks in YQHX preparations. These data provided valuable information, on the molecular weights and structure of the constituents. (DOCX 27 kb)

In the past, the optimal first-line treatment of advanced non-small cell lung cancer (NSCLC) patients with wild-type for epidermal growth factor receptor (wt-EGFR) has been platinum-based chemotherapy doublets (1,2). More recently, clinicians have adopted immunotherapy with check-point inhibitors alone, in PD-L1 strong positive, or combined with chemotherapy independently to PD-L1 expression (3).

Tobacco smoke (TS) is a major causative agent to lead to chronic bronchitis (CB). However the mechanisms of CB induced by TS are unclear. In this report, rats were exposed to different concentrations of TS and the metabolic features of CB were characterized by using a nontargeted metabolic profiling method based on liquid chromatography-mass spectrometry (LC-MS) to detect the altered metabolic patterns in serum from CB rats and investigate the mechanisms of CB. 11 potential biomarkers were identified in serum of rats. Among them, the levels of lysophosphatidylethanolamine (18:1), lysophosphatidic acid (18:1), lysophosphatidylethanolamine (18:0), lysophosphatidylethanolamine (16:0), lysophosphatidylethanolamine (20:4), docosahexaenoic acid, 5-hydroxyindoleacetic acid and 5'-carboxy-γ-tocopherol were higher in TS group compared to control group. Conversely, the levels of 4-imidazolone-5-propionic acid, 12-hydroxyeicosatetraenoic acid and uridine were lower in TS group. The results indicated that the mechanism of CB was related to amino acid metabolism and lipid metabolism, particularly lipid metabolism. In addition, lysophosphatidylethanolamines were proved to be important mediators, which could be used as biomarkers to diagnose CB. These results also suggested that metabolomics was suitable for diagnosing CB and elucidating the possible metabolic pathways of TS-induced CB.

Objective: The present work aimed to retrospectively assess the outcomes associated with decortication by video-assisted thoracic surgery (VATS) in patients with tuberculous empyema. Methods: Patients (n = 274) who underwent decortication by VATS for surgical management of pleural empyema between January 2000 to 2010 were included. Pre-, intra-, and post-operative characteristics were observed for all patients, which were followed up for 12 months to evaluate surgical outcomes such as postoperative complications and disease recurrence. Results: No patients required conversion to thoracotomy, and no death or postoperative bleeding was reported. The mean operation time was 104.5 ± 20.4 min, with 271.5 ± 41.3 ml intraoperative blood loss and median length of hospital stay of 7.2 ± 3.4 days. Of the 274 patients, 262 were followed up for 12 months; 26 (9.9%) patients showed complications, including incomplete lung re-expansion (11 patients) and persistent air leak (6 patients). While early disease recurrence was observed in 3 (1.1%) patients after surgery, late recurrence was reported for 6 (2.3%) individuals. Interestingly, the complication rate was much higher in patients with chronic empyema (15/34, 44.1%) than in subjects with acute empyema (11/228, 4.8%). Conclusions: Decortication by VATS decreases postsurgical complications, and results in decreased disease recurrence. This study demonstrated improved outcomes by decortication by VATS, even in patients with stage III tuberculous empyema.

Argonaute (AGO) family proteins are conserved key components of small RNA-induced silencing pathways. In the RNA-directed DNA methylation (RdDM) pathway in Arabidopsis, AGO6 is generally considered to be redundant with AGO4. In this report, our comprehensive, genomewide analyses of AGO4- and AGO6-dependent DNA methylation revealed that redundancy is unexpectedly negligible in the genetic interactions between AGO4 and AGO6. Immunofluorescence revealed that AGO4 and AGO6 differ in their subnuclear co-localization with RNA polymerases required for RdDM. Pol II and AGO6 are absent from perinucleolar foci, where Pol V and AGO4 are co-localized. In the nucleoplasm, AGO4 displays a strong co-localization with Pol II, whereas AGO6 co-localizes with Pol V. These patterns suggest that RdDM is mediated by distinct, spatially regulated combinations of AGO proteins and RNA polymerases. Consistently, Pol II physically interacts with AGO4 but not AGO6, and the levels of Pol V-dependent scaffold RNAs and Pol V chromatin occupancy are strongly correlated with AGO6 but not AGO4. Our results suggest that AGO4 and AGO6 mainly act sequentially in mediating small RNA-directed DNA methylation.

Introduction Surgical ablation is a generally established treatment for patients with atrial fibrillation undergoing concomitant cardiac surgery. Left atrial (LA) lesion set for ablation is a simplified procedure suggested to reduce the surgery time and morbidity after procedure. The present meta-analysis aims to explore the outcomes of left atrial lesion set versus no ablative treatment in patients with AF undergoing cardiac surgery. Methods A literature research was performed in six database from their inception to July 2017, identifying all relevant randomized controlled trials (RCTs) comparing left atrial lesion set versus no ablative treatment in AF patient undergoing cardiac surgery. Data were extracted and analyzed according to predefined clinical endpoints. Results Eleven relevant RCTs were included for analysis in the present study. The prevalence of sinus rhythm in ablation group was significantly higher at discharge, 6-month and 1-year follow-up period. The morbidity including 30 day mortality, late all-cause mortality, reoperation for bleeding, permanent pacemaker implantation and neurological events were of no significant difference between two groups. Conclusions The result of our meta-analysis demonstrates that left atrial lesion set is an effective and safe surgical ablation strategy for AF patients undergoing concomitant cardiac surgery.

Background The causal link between tobacco smoke exposure (TSE) and numerous severe respiratory system diseases (RSD), including chronic bronchitis, chronic obstructive pulmonary disease, and lung cancer, is well established. However, the pathogenesis of TSE-induced RSD remains incompletely understood. This research aims to detect the pathogenetic mechanisms and potential therapeutic targets of TSE-induced RSD. Methods This study employed TSE model which rats were exposed to a concentration of 60% tobacco smoke in a toxicant exposure system for four weeks. Tandem mass tags (TMT) labeled quantitative proteomics combined with off-line high pH reversed-phase fractionation, and nano-liquid chromatography-mass spectrometry method (off-line high pH RPF-nano-LC–MS/MS) were adopted to detect differentially expressed proteins (DEPs) in the lung tissues of the TSE model rats and to compare them with those in control. The accuracy of the results was verified by western blot. Results Compared with the control group, 33 proteins in the TSE model group's lung tissues showed significant differential expression. Analysis based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways indicated that, several biological pathways, such as the steroid biosynthesis pathway, were involved and played significant roles in the pathogenesis of the experimental group's TSE. Conclusions These findings make a crucial contribution to the search for a comprehensive understanding of TSE-induced RSD's pathogenesis, and furthermore provide guidance for the diagnosis and treatment of TSE-induced RSD.

Pollen development is an important and complex biological process in the sexual reproduction of flowering plants. Although the cytological characteristics of pollen development are well defined, the regulation of its early stages remains largely unknown. In the present study, miRNAs were explored in the early development of broccoli (Brassica oleracea var. italica) pollen. A total of 333 known miRNAs that originated from 235 miRNA families were detected. Fifty-five novel miRNA candidates were identified. Sixty of the 333 known miRNAs and 49 of the 55 predicted novel miRNAs exhibited significantly differential expression profiling in the three distinct developmental stages of broccoli pollen. Among these differentially expressed miRNAs, miRNAs that would be involved in the developmental phase transition from uninucleate microspores to binucleate pollen grains or from binucleate to trinucleate pollen grains were identified. miRNAs that showed significantly enriched expression in a specific early stage of broccoli pollen development were also observed. In addition, 552 targets for 127 known miRNAs and 69 targets for 40 predicted novel miRNAs were bioinformatically identified. Functional annotation and GO (Gene Ontology) analysis indicated that the putative miRNA targets showed significant enrichment in GO terms that were related to plant organ formation and morphogenesis. Some of enriched GO terms were detected for the targets directly involved in plant male reproduction development. These findings provided new insights into the functions of miRNA-mediated regulatory networks in broccoli pollen development.

Dehydration responsive element binding proteins are transcription factors of the plant-specific AP2 family, many of which contribute to abiotic stress responses in several plant species. We investigated the possibility of increasing drought tolerance in the traditional Chinese medicinal herb, Salvia miltiorrhiza, through modulating the transcriptional regulation of AtDREB1C in transgenic plants under the control of a constitutive (35S) or drought-inducible (RD29A) promoter. AtDREB1C transgenic S. miltiorrhiza plants showed increased survival under severe drought conditions compared to the non-transgenic wild-type (WT) control. However, transgenic plants with constitutive overexpression of AtDREB1C showed considerable dwarfing relative to WT. Physiological tests suggested that the higher chlorophyll content, photosynthetic capacity, and superoxide dismutase, peroxidase, and catalase activity in the transgenic plants enhanced plant drought stress resistance compared to WT. Transcriptome analysis of S. miltiorrhiza following drought stress identified a number of differentially expressed genes (DEGs) between the AtDREB1C transgenic lines and WT. These DEGs are involved in photosynthesis, plant hormone signal transduction, phenylpropanoid biosynthesis, ribosome, starch and sucrose metabolism, and other metabolic pathways. The modified pathways involved in plant hormone signaling are thought to be one of the main causes of the increased drought tolerance of AtDREB1C transgenic S. miltiorrhiza plants.

The present study developed a systematic method for the degradation products and dynamics of valtrate in artificial intestinal fluid and gastric fluid. HPLC-electrospray tandem mass spectrometry (HPLC-ESI-MSn) was applied to separate the degradation products of valtrate and identify structures of the main degradation products. Samples were separated on an Agilent Extend-Cl8 by gradient elution using acetonitrile and water–acetic acid (0.25%, v/v). Analysis was performed in positive ion mode with 3000 V of capillary voltage, 40 psi of atomization pressure, 10 L min−1 dry nitrogen gas velocity, and m/z 100–1000 of scan range. Six degradation products were all identified to own the stem-nucleus structures of baldrinals. Two of them were identified through the reference substances. An HPLC method with isocratic elution using acetonitrile and water (68:32, v/v) at 254 nm was established to determine content change of valtrate in artificial intestinal fluid and gastric fluid at different time points. The method...

DNA methylation is one of the most important epigenetic modifications involved in the development and differentiation in plants. Hypocotyl and cotyledon are the two major tissues of cauliflower (Brassica oleracea L. var. botrytis) seedlings. Both tissues show significantly different tissue specificity and regenerative abilities in vitro. However, the characteristics of DNA methylation modification and its roles in regulating the organ development in cauliflower remain largely unknown. In the present study, the DNA methylation status between the hypocotyl and cotyledon of cauliflower seedlings were analyzed. The results indicated that although the hypocotyl and cotyledon of cauliflower seedlings share the same genome, the genomic DNA methylation levels and patterns at CCGG sites were different. Compared with the cotyledon, the hypocotyl showed higher DNA methylation level, and more loci showing methylation pattern adjustments were also discovered. Twelve loci with changes of DNA methylation patterns were further explored. The quantitative expression analysis indicated that eight out of twelve sequenced fragments showed differential expression between the hypocotyl and cotyledon, of which the expression of six sequences was identified to be negative correlation with their DNA methylation status. In addition, three main DNA methyltransferase genes MET1, CMT3 and DRM were first explored in cauliflower. The results indicated that the expression of these three genes was closely associated with the different DNA methylation status in the hypocotyl and cotyledon. These findings provided more information to further explore the roles of DNA methylation modification in tissue differentiation and development of cauliflower.

Valtrate is a principle compound isolated fromValeriana jatamansiJones, which is a Traditional Chinese Medicine used to treat various mood disorders. The aim of the present study was to investigate the anxiolytic effects of valtrate in rats. The animals were orally administered valtrate (5, 10, and 20 g/kg daily) for 10 days and exposed to open field test (OFT) and elevated plus-maze (EPM). Then the corticosterone levels in the rat serum were measured by enzyme-linked immunosorbent assay (ELISA). The valtrate (10 mg/kg, p.o.) exhibited the anxiolytic effect in rats by increasing the time and entry percentage into the open arms in the EPM and the number of central entries in the OFT. Valtrate (10 mg/kg, p.o.) significantly reduced the corticosterone level in the rat serum. Taken together, these results suggest that the valtrate has anxiolytic activity in behavioral models that might be mediated via the function of hypothalamus-pituitary-adrenal axis.

Introduction An atrial septal aneurysm (ASA) is a rare congenital cardiac deformity characterized by interatrial septum protruding into atria forming a saccular structure. Patient concerns In our case, a 42-year-old female patient presented to our hospital complained of palpitation. Diagnosis Transthoracic echocardiography detected a 3.4 × 3.4 cm circular mass attached to the interatrial septum in right atrium complicated with a 6 mm secundum atrial septal defects (ASD). Interventions The patient received a cardiopulmonary bypass surgery to remove the mass and close the ASD. Outcomes The mass turned out to be an organized thrombus with calcium deposition and fibrinoid necrosis. Conclusion ASA is a potential location of atrial thrombus because of the stagnation of blood. Systemic embolism events are the main complications of ASA. Surgery or anticoagulation is both recommended in patients with ASA with thrombus.

In Larix and in some crops, heterosis is prevalent and has been widely used in breeding to produce excellent varieties. However, the molecular basis of heterosis in Larix remains ambiguous. LaAP2L1, a member of the AP2/EREBP transcription factor family, has been suggested to be involved in heterosis in Larix hybrids. Here, the function and regulation of LaAP2L1 were further explored. Overexpression of LaAP2L1 led to markedly enlarged organs and heterosis-like traits in Arabidopsis. Fresh weight of leaves was almost twice as great as in vector controls. Likewise, seed yield of 35S::LaAP2L1 individual plants was >200% greater than that of control plants. The enlarged organs and heterosis-like traits displayed by 35S::LaAP2L1 plants were mainly due to enhanced cell proliferation and prolonged growth duration. At the molecular level, LaAP2L1 upregulated the expression of ANT, EBP1, and CycD3;1 and inhibited the expression of ARGOS in 35S::LaAP2L1 plants, suggesting an important molecular role of LaAP2L1 in regulating plant organ development. These findings provide new insights into the formation of heterosis in woody plants and suggest that LaAP2L1 has potential applications in breeding high-yielding crops and energy plants. In addition, 50 AP2/EREBP transcription factors, including LaAP2L1, in Larix were identified by transcriptome sequencing, and phylogenetic analysis was conducted. This provided information that will be important in further revealing the functions of these transcription factors.

Esophageal squamous cell carcinoma (ESCC) is one of the most common tumors worldwide, with a high malignant degree and poor prognosis. The present study aims to investigate the relationship between pyruvate kinase M2 (PKM2) expression and the prognosis of patients with ESCC. The expression of PKM2 in 86 cases of esophageal carcinoma tissues was tested using immunohistochemistry. The relationship between PKM2 expression and clinical pathological parameters, and their effects on the prognosis of patients with ESCC were analyzed. The expression levels of PKM2 in both cytoplasm and nucleus of ESCC tissues were significantly higher than those in paracancerous tissues (P = 6.73 × 10−9 and 4.32 × 10−6, respectively). The Kaplan–Meier analysis showed that nuclear PKM2 expression was closely related to the survival of patients with ESCC (P = 0.005). Patients with high PKM2 expression in the nucleus had significantly shorter survival times than those with low PKM2 expression in the nucleus (hazard ratio for death, 2.358; 95% confidence interval, 1.156–4.812; P = 0.018). No other significant difference was found between PMK2 expression and clinico-pathological features of ESCC patients (all P > 0.05). In conclusion, high PKM2 expression in the nucleus is essential in the pathogenic process of ESCC and may be used to predict the prognosis of patients with ESCC.

Heterosis is prevalent in Larix kaempferi, but the molecular mechanism underlying heterosis in this species has not yet been elucidated. We have explored the relationships among heterosis and DNA sequence variations, changes in the DNA methylation status, and gene expression in a set of Larix kaempferi intraspecific parental lines and their reciprocal hybrids. The results indicate that the two intraspecific parental lines have highly similar genome structures and that the crosses did not produce substantial changes in the genomic DNA of the hybrid progeny. In contrast, there was a significant difference in the DNA methylation status between the two parental lines. The genomic DNA methylation level in the heterotic hybrids (26.47 %) was significantly lower than the midparent value (MPV) (33.80 %), indicating that significant extensive demethylation had occurred. However, the non-heterotic hybrids displayed a DNA methylation level (33.59 %) that was close to the MPV, and they contained similar numbers of methylation and demethylation events. The correlation analysis revealed that the increased gene expression in the heterotic hybrids was associated with its overall low genomic DNA methylation level and dynamic DNA methylation patterns. Twenty fragments with adjusted DNA methylation patterns were then cloned and the transcript level analyzed quantitatively. The results further indicated that the expressions of most but not all of the 12 possible genes were negatively correlated to their respective DNA methylation status. Based on these data, we suggest that adjustments in DNA methylation status may play critical roles in the formation of heterosis produced by the cross between two Larix kaempferi intraspecific parental lines.

Pollen development is an important and complex biological process in the sexual reproduction of flowering plants. Although the cytological characteristics of pollen development are well defined, the regulation of its early stages remains largely unknown. In the present study, miRNAs were explored in the early development of broccoli (Brassica oleracea var. italica) pollen. A total of 333 known miRNAs that originated from 235 miRNA families were detected. Fifty-five novel miRNA candidates were identified. Sixty of the 333 known miRNAs and 49 of the 55 predicted novel miRNAs exhibited significantly differential expression profiling in the three distinct developmental stages of broccoli pollen. Among these differentially expressed miRNAs, miRNAs that would be involved in the developmental phase transition from uninucleate microspores to binucleate pollen grains or from binucleate to trinucleate pollen grains were identified. miRNAs that showed significantly enriched expression in a specific early stage of broccoli pollen development were also observed. In addition, 552 targets for 127 known miRNAs and 69 targets for 40 predicted novel miRNAs were bioinformatically identified. Functional annotation and GO (Gene Ontology) analysis indicated that the putative miRNA targets showed significant enrichment in GO terms that were related to plant organ formation and morphogenesis. Some of enriched GO terms were detected for the targets directly involved in plant male reproduction development. These findings provided new insights into the functions of miRNA-mediated regulatory networks in broccoli pollen development.

Heterosis has been widely explored in Larix breeding for more than a century, but the molecular mechanisms underlying this phenomenon remain elusive. In the present study, the genome-wide transcript profiles from two Larix genotypes and their reciprocal hybrids were analyzed using Arabidopsis 70-mer oligonucleotide microarrays. Despite sharing the same two parental lines, one of the hybrids showed obvious heterosis, while the other did not. In total, 1,171 genes were differentially expressed between the heterotic hybrid and its parents, of which 133 genes were nonadditive expression. The number of differentially expressed genes between the non-heterotic hybrid and the parents was 939, but only 54 of these genes were nonadditive expression. Further, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses indicated that most of these differentially expressed genes in the heterotic hybrid were associated with several important biological functions such as physiological processes, responses to stimulus, and starch and sucrose metabolism. The reliability of the microarray data was further validated by the Real-time quantitative RT-PCR. A high Pearson linear correlation coefficient value was detected (r = 0.759, P

The majority of verified plant disease resistance genes (R genes) isolated to date was of the nucleotide binding site-leucine rich repeat (NBS-LRR) class. The conservation between different NBS-LRR R genes opens the avenue for the use of PCR based strategies in isolating and cloning other R gene family members or analogs (resistance gene analogue, RGA) using degenerate primers for these conserved regions. In this study, to better understand the R gene in European aspen (Populus tremula), a perennial tree, we used degenerate primers to amplify RGA sequences from European aspen. Cloning and sequence characterization identified 37 European aspen RGAs, which could be phylogenetically classified into seven subfamilies. Deduced amino acid sequences of European aspen RGAs showed strong identity, ranging from 30.41 to 46.63%, to toll interleukin receptor (TIR) R gene subfamily. BLAST searches with reference to the genomic sequence of P. trichocarpa found 209 highly homologous regions distributed in 28 genomic loci, suggesting the abundance and divergence of NBS-encoding R genes in European aspen genome. Although, numerous studies have reported that plant R genes are under diversifying selection for specificity to evolving pathogens, non-synonymous to synonymous nucleotide substitution (dN/dS) ratio were

Amplified fragment length polymorphism (AFLP) was used in genetic fingerprinting of 22 elite Chinese Populus deltoides and Populus × canadensis accessions. The results indicated that each of the nine AFLP primer pairs selected generated fingerprint profiles that were unique to each of the accessions. Therefore, each accession was definitively identified by any of the nine primer pairs. Furthermore, the inter-accession genetic relationships inferred based on 461 polymorphic fragments from the nine AFLP primer pairs were largely consistent with phylogenetic relationships based on morphologic traits. Bootstrap analysis showed that three AFLP primer pairs were required to obtain genetic similarity values with a maximum CV of 10% while 10 AFLP primer pairs could give a maximum CV of 5%. Thus, AFLP can readily be applied for a rapid and accurate evaluation of the degree of similarity between poplar cultivars. In this study, the number of AFLP fragments used was sufficient to establish a reliable estimate of genetic similarity among accessions, with a maximum CV of 5.12%. Therefore, the information on the genetic relationships among the poplar accessions generated in this study in connection with knowledge on agronomic traits may have an impact on poplar breeding and planting in China.

In this study, bulked segregant analysis (BSA) was used on Larix leptolepis × Larix olgensis hybrids to identify a random amplified polymorphic DNA (RAPD) marker associated with high rooting ability in larch. Two DNA bulks: H (high rooting ability) bulk and L (low rooting ability) bulk were constructed according to the rooting percentages of the stock plants. Among the 328 primers, only S356 could amplify a specific band, named S356445, which only existed in the H bulk and was further confirmed following selective genotyping of individual hybrids. Grounded on the border sequences, S356445 was converted to a sequence characterized amplified region (SCAR) marker, HRL445, which can be useful in marker-assisted selection (MAS) to screen for larch with high rooting ability. All the results strongly indicated that S356445 and HRL445 were closely associated with high rooting ability in larch.

In this study, seven cDNA fragments of genes, differentially expressed in ovaries after pollination in Cymbidium hybridium, were identified and characterized by mRNA differential display reverse transcription polymerase chain reaction (DDRT-PCR). Four (CDD-313, CDD-272, CDD-265, CDD-243) among these seven cDNA fragments showed no significant homology with ESTs or genes in the databases of NCBI; another three (CDD-193, CDD-218, CDD-470) showed significant homologies with sequences encoding components of an ABC-type transporter, a GTPase and 40S ribosomal S3 proteins (RPS3), respectively. The differential expression patterns of them were confirmed by reverse Northern dot blot analysis. More interestingly, CDD-470 appeared to be present and highly expressed in the pollinated ovaries and encoded a new factor of RPS3 participating in cell growth and proliferation. We deduced that this 40s ribosomal S3 like protein was involved in ovary development of orchids.

Accurate identification of varieties (clones) and knowledge of their genetic relationships are essential for poplar breeding and variety management. In this study, 21 elite poplar varieties of Tacamahaca and Aigeiros in China were fingerprinted using amplified fragment length polymorphism (AFLP) markers. Four AFLP primer pairs developed generated totally 181 AFLP polymorphic fragments, and in particular, each primer pair generated fingerprint profiles specific to each of the tested varieties. The genetic relationships among the varieties were evaluated by dendrograms and multidimensional scaling (MDS). The results showed that the tested poplar can be classified into five groups, and indicated the clear separation of varieties of different sections of poplar and the primary distinction between native and exotic poplar varieties. This study indicated that tested poplar varieties could be identified by their fingerprint profiles and that genetic relationships deduced from the study were consistent with their genealogy. In addition, our results demonstrated that AFLP could be used to construct DNA fingerprints of polar clones at a large-scale level and to determine genetic relationships of poplar varieties.

Multiple stress response genes are controlled by transcription factors in a coordinated manner; therefore, these factors can be used for molecular plant breeding. CBF1/DREB1B, a known stress-inducible gene, was isolated from Arabidopsis thaliana and introduced into Salvia miltiorrhiza under the control of the CaMV35S or RD29A promoter. Under drought stress, relative water content, chlorophyll content, and the net photosynthetic rate were observed to be higher in the transgenic lines than in the wild type (WT). Moreover, O2(-) and H2O2 accumulation was observed to be lower in the transgenic lines. Additional analyses revealed that the AtDREB1B transgenic plants generally displayed lesser malondialdehyde (MDA) but higher superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities than the WT under drought stress. Quantitative real-time polymerase chain reaction of a subset of genes involved in photosynthesis, stress response, carbohydrate metabolism, and cell protection further verified that AtDREB1B could enhance tolerance to drought by activating different downstream DREB/CBF genes in the transgenic plants. Furthermore, no growth inhibition was detected in transgenic S. miltiorrhiza plants that expressed AtDREB1B driven by either the constitutive CaMV35S promoter or the stress-inducible RD29A promoter. Together, these results suggest that AtDREB1B is a good candidate gene for increasing drought tolerance in transgenic S. miltiorrhiza.

Tobacco smoke (TS) is a major causative agent to lead to chronic bronchitis (CB). However the mechanisms of CB induced by TS are unclear. In this report, rats were exposed to different concentrations of TS and the metabolic features of CB were characterized by using a nontargeted metabolic profiling method based on liquid chromatography-mass spectrometry (LC-MS) to detect the altered metabolic patterns in serum from CB rats and investigate the mechanisms of CB. 11 potential biomarkers were identified in serum of rats. Among them, the levels of lysophosphatidylethanolamine (18:1), lysophosphatidic acid (18:1), lysophosphatidylethanolamine (18:0), lysophosphatidylethanolamine (16:0), lysophosphatidylethanolamine (20:4), docosahexaenoic acid, 5-hydroxyindoleacetic acid and 5'-carboxy-γ-tocopherol were higher in TS group compared to control group. Conversely, the levels of 4-imidazolone-5-propionic acid, 12-hydroxyeicosatetraenoic acid and uridine were lower in TS group. The results indicated that the mechanism of CB was related to amino acid metabolism and lipid metabolism, particularly lipid metabolism. In addition, lysophosphatidylethanolamines were proved to be important mediators, which could be used as biomarkers to diagnose CB. These results also suggested that metabolomics was suitable for diagnosing CB and elucidating the possible metabolic pathways of TS-induced CB.

The Inter-Simple Sequence Repeat (ISSR) was used in this study for genetic fingerprinting and identification of 28 important Populus L. (poplar) cultivars (varieties/ clones), and determination of the genetic relationships among these cultivars. These 28 cultivars belonged to sections Aigeiros, Tacahamaca, Leuce, Turanga, and hybrids between sections Aigeiros and Tacahamaca. Out of 27 ISSR primers tested, eight primers generated clear multiplex profiles. The best three primers produced 154 easily detectable fragments, 129 (84%) of which were polymorphic among the cultivars. Each of these 3 primers produced fingerprint profiles unique to each of the accessions studied, and thus could be solely used for their identification. Twenty-five markers, unique to 10 of the cultivars studied, were detected. These markers may be converted into cultivar-specific probes for identification purposes. Genetic relationships among the cultivars were evaluated by generating a similarity matrix based on the simple matching coefficient and the unweighted pair group method with arithmetic average (UPGMA) dendrogram. The results showed a clear-cut separation of cultivars among different sections of poplar, and were in agreement with the genealogy of the sampled cultivars. The present study shows that ISSR markers could generate abundant polymorphism, are reproducible, and are quick for characterization of poplar cultivars. In the future, the markers used in this study, in combination with other molecular techniques, could provide a useful panel of ISSR markers for largescale DNA fingerprinting of poplar cultivars and determination of the genetic relationships among these cultivars.

Argonaute (AGO) family proteins are conserved key components of small RNA-induced silencing pathways. In the RNA-directed DNA methylation (RdDM) pathway in Arabidopsis, AGO6 is generally considered to be redundant with AGO4. In this report, our comprehensive, genomewide analyses of AGO4- and AGO6-dependent DNA methylation revealed that redundancy is unexpectedly negligible in the genetic interactions between AGO4 and AGO6. Immunofluorescence revealed that AGO4 and AGO6 differ in their subnuclear co-localization with RNA polymerases required for RdDM. Pol II and AGO6 are absent from perinucleolar foci, where Pol V and AGO4 are co-localized. In the nucleoplasm, AGO4 displays a strong co-localization with Pol II, whereas AGO6 co-localizes with Pol V. These patterns suggest that RdDM is mediated by distinct, spatially regulated combinations of AGO proteins and RNA polymerases. Consistently, Pol II physically interacts with AGO4 but not AGO6, and the levels of Pol V-dependent scaffold RNAs and Pol V chromatin occupancy are strongly correlated with AGO6 but not AGO4. Our results suggest that AGO4 and AGO6 mainly act sequentially in mediating small RNA-directed DNA methylation.

Polyploidization is known to accompany altered DNA methylation in higher plants, which plays an important role in gene expression regulation and maintaining genome stability. While the characteristics of DNA methylation in different polyploid plants are still to be elucidated; here, status of genomic DNA methylation in a series of diploid, triploid, and tetraploid annual herbaceous plants (watermelon and Salvia) and woody perennials (pear, Poplar, and loquat) were explored by methylation-specific amplified polymorphism analysis. The results indicated that levels of DNA methylation in triploid watermelon and Salvia were lower than their diploid parents. In triploid Poplar and pear, higher levels of DNA methylation were detected, and no significant difference was observed between triploid and tetraploid in all tested materials. Further data analysis suggested that about half of the total detected sites underwent changes of DNA methylation patterns in triploid watermelons and Salvia, as well as an obvious trend towards demethylation. However, the changes of DNA methylation patterns in three triploid woody perennials were only 17.54–33.40%. This implied that the characteristics of DNA methylation are significantly different during the polyploidization of different plant species. Furthermore, the results suggested that the level of DNA methylation was nonlinearly related to the ploidy level, and triploid plants displayed more interesting DNA methylation status. The characteristics and possible functions of DNA methylation in different ploidy series are further discussed.

MicroRNAs (miRNAs) are a class of small endogenous, non-coding RNAs that have key regulatory functions in plant growth, development, and other biological processes. Hypocotyl and cotyledon are the two major tissues of cauliflower (Brassica oleracea L. var. botrytis) seedlings. Tissue culture experiments have indicated that the regenerative abilities of these two tissues are significantly different. However, the characterization of miRNAs and their roles in regulating organ development in cauliflower remain unexplored. In the present study, two small RNA libraries were sequenced by Solexa sequencing technology. 99 known miRNAs belonging to 28 miRNA families were identified, in which 6 miRNA families were detected only in Brassicaceae. A total of 162 new miRNA sequences with single nucleotide substitutions corresponding to the known miRNAs, and 32 potentially novel miRNAs were also first discovered. Comparative analysis indicated that 42 of 99 known miRNAs and 17 of 32 novel miRNAs exhibited significantly differential expression between hypocotyl and cotyledon, and the differential expression of several miRNAs was further validated by stem-loop RT-PCR. In addition, 235 targets for 89 known miRNAs and 198 targets for 24 novel miRNAs were predicted, and their functions were further discussed. The expression patterns of several representative targets were also confirmed by qRT-PCR analysis. The results identified that the transcriptional expression patterns of miRNAs were negatively correlated with their targets. These findings gave new insights into the characteristics of miRNAs in cauliflower, and provided important clues to elucidate the roles of miRNAs in the tissue differentiation and development of cauliflower.

In 2001, when appraisal and development work began at Daan field, the MI Energy-PetroChina partnership was faced with the commercial necessity to optimize production. Given the relatively low oil prices that prevailed then, the Daan field project was considered marginally commercial. It was imperative to define a cost-effective and technologically strong field development program. The main geologic risk factor was reservoir sandstone distribution, and a new model was constructed to predict distribution of the primary reservoir objective FuYang sandstones. Consistent with the structural evolution of the Daan area, the FuYang reservoir is focused on a series of north-northeast to northeast-trending depocenters controlled by down-to-the-northwest normal faults parallel and genetically related to the Daan fault system. The Fuyang reservoir sandstones and siltstones are interpreted to have been deposited in fluvial and marginal lacustrine environments associated with small- to medium-size lake systems, which developed in the north-northeast to northeast-trending depocenters. Based on core studies and observed modern analogs, the main FuYang reservoir facies comprised lake-delta mouth bar and fluvial channel-bar deposits. The FuYang reservoir is best developed in the north-northeast to northeast-trending sweet spots, which correspond to the structurally controlled depocenters. Although the FuYang is also oil productive at Daan outside these sweet spot areas, well productivity is lower because of reduced reservoir sandstone development. The development plan for the Daan field called for wells to be drilled in a consistent grid pattern (175 350 m [574 1148 ft]), but the order of drilling has a significant impact on the commerciality of the project, and the reservoir distribution model is key to defining the drilling order. Appraisal wells are drilled continuously in an effort to further refine the reservoir model and identify FuYang sweet spot areas. At the same time, development well drilling proceeds in priority order, focusing first on the best quality FuYang sweet spot areas. This systematic drilling approach has lowered reservoir risk and improved early well productivity by as much as 40 to 50%, thereby significantly enhancing the project's profitability. Understanding of the FuYang reservoir distribution has also contributed to a large increase in the estimate of oil in place for the Daan field.

The aim of the study was the explain the mechanism related to therapeutic effects of Uremic Clearance Granules (Niaoduqing Keli in Chinese) on adenine-induced Chronic Renal Failure in rats. Thirty 8-week-old male Wistar rats were selected and randomly divided in to 3 groups: Normal Control Group (NCG)consisted of 10 rats, Chronic Renal Failure Pathological Control Group (PCG) 10 rats, and Uremic Clearance Granules Treatment Group (UCG) 10 rats. Each rat in PCG and UCG was fed with adenine-enriched diets, containing 10 g adenine per kg food for 6 weeks. After fed with adenine, each rat in UCG was administered orally with 2 ml solution of Uremic Clearance Granules for 6 weeks. The concentration of Uremic Clearance Granules solution was 0.42 g/ml which was 10 times of human. On days 42 and 84, the serum levels of creatinine, Blood Urea Nitrogen and homocysteine were determined. The methylation of TGFbeta1 promoter was tested by methylation-specific PCR. TGF-beta1 mRNA and protein expression in rat renal cortex were analyzed by real-time RT-PCR and Immunohistochemistry. (1) Experimented on model of Chronic Renal Failure in rats, the preparation was proved to be able to reduce serum creatinine, Blood Urea Nitrogen, and homocysteine (p

DNA demethylation mediated by the DNA glycosylase ROS1 helps determine genomic DNA methylation patterns and protects active genes from being silenced. However, little is known about the mechanism of regulation of ROS1 enzymatic activity. Using a forward genetic screen, we identified an anti-silencing (ASI) factor, ASI3, the dysfunction of which causes transgene promoter hyper-methylation and silencing. Map-based cloning identified ASI3 as MET18, a component of the cytosolic iron-sulfur cluster assembly (CIA) pathway. Mutation in MET18 leads to hyper-methylation at thousands of genomic loci, the majority of which overlap with hypermethylated loci identified in ros1 and ros1dml2dml3 mutants. Affinity purification followed by mass spectrometry indicated that ROS1 physically associates with MET18 and other CIA components. Yeast two-hybrid and split luciferase assays showed that ROS1 can directly interact with MET18 and another CIA component, AE7. Site-directed mutagenesis of ROS1 indicated that the conserved iron-sulfur motif is indispensable for ROS1 enzymatic activity. Our results suggest that ROS1-mediated active DNA demethylation requires MET18-dependent transfer of the iron-sulfur cluster, highlighting an important role of the CIA pathway in epigenetic regulation., Author Summary DNA cytosine methylation is a major epigenetic mark that confers transcriptional regulation. Active removal of DNA methylation is important for plants and mammals during development and in responses to various stress conditions. In the model plant species Arabidopsis thaliana, active DNA demethylation depends on a family of 5-methylcytosine DNA glycosylases/demethylases including ROS1, DME, and others. While the epigenetic function of this demethylase family is well-known, little is known about how their enzymatic activities may be regulated. In this report, we carried out a forward genetic screen for anti-silencing factors and identified MET18, a conserved component of cytosolic iron-sulfur cluster assembly (CIA) pathway in eukaryotes, as being required for the ROS1-dependent active DNA demethylation. Dysfunction of MET18 causes DNA hyper-methylation at thousands of genomic loci where DNA methylation is pruned by ROS1. In addition, ROS1 physically interacts with MET18 and other CIA pathway components; while a conserved iron-sulfur-binding motif is indispensable for ROS1 enzyme activity. Our results suggested that MET18 affects DNA demethylation by influencing ROS1 enzymatic activity via direct interaction with the iron-sulfur-binding motif of ROS1, highlighting a direct connection between iron-sulfur cluster assembly and active DNA demethylation.

The control of autonomous underwater vehicles (AUV) presents a number of unique and formidable challenges. AUV dynamics is highly nonlinear, coupled, time varying and subject to hydrodynamic uncertainties and external disturbances such as current. In this paper, we presented a new type of controller for AUV motion control based on fuzzy cerebella model articulation controller (FCMAC). A nonlinear six degree-of-freedom equation of AUV motion was derived. The application of the designed FCMAC controller to AUV motion control was discussed and the simulation results shown that this algorithm possessed better dynamic performance and robustness than conventional PID strategy.