Autophagy-mediated degradation of integumentary tapetum is critical for embryo pattern formation.

Autophagy modulates the degradation and recycling of intracellular materials and contributes to male gametophyte development and male fertility in plants. However, whether autophagy participates in seed development remains largely unknown. Here, we demonstrate that autophagy is crucial for timely programmed cell death (PCD) in the integumentary tapetum, the counterpart of anther tapetum, influencing embryo pattern formation and seed viability. Inhibition of autophagy resulted in delayed PCD of the integumentary tapetum and defects in embryo patterning. Cell-type-specific restoration of autophagic activities revealed that the integumentary tapetum plays a non-autonomous role in embryo patterning. Furthermore, high-throughput, comprehensive lipidomic analyzes uncovered an unexpected seed-developmental-stage-dependent role of autophagy in seed lipid metabolism: it contributes to triacylglycerol degradation before fertilization and to triacylglycerol biosynthesis after fertilization. This study highlights the critical role of autophagy in regulating timely integumentary tapetum PCD and reveals its significance in seed lipid metabolism and viability.

Analysis of plasma free amino acids in diabetic rat and the intervention of Ginkgo biloba leaves extract using hydrophilic interaction liquid chromatography coupled with tandem mass-spectrometry.

Amino acids (AAs) are important metabolites that are related with diabetes. However, their roles in the initiation and development of diabetes mellitus (DM), especially in the treatment of Ginkgo biloba leaves extract (GBE) have not been fully explored. Thus, we investigated the roles that AAs played in the progression and GBE supplementation of DM rat induced by streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution, a normal control group treated with GBE, a DM group treated with drug-free solution, and DM group treated with GBE; and maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with the optimized hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method. A total of 17 AAs with differential levels were monitored to indicate dysfunction of AAs metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in seven AAs including leucine, isoleucine, tyrosine, glutamic acid, asparagines, lysine and alanine in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on AAs metabolism. The improvement of those AAs metabolism disorders may play a considerable role in the treatment of GBE on the occurrence and development of DM. Those findings potentially promote the understanding of the pathogenic progression of DM and reveal the therapeutic mechanism of GBE against DM.

Autophagy-mediated compartmental cytoplasmic deletion is essential for tobacco pollen germination and male fertility.

In plants, macroautophagy/autophagy has mainly been associated with stress-related processes but how it impacts normal physiological and developmental processes remains largely unexplored. Pollen germination is the critical first step toward fertilization in flowering plants. It is metabolically demanding and relies on high levels of cytoplasmic reorganization activities to support a dramatic morphological transformation that underlies the development of a pollen tube as the conduit to deliver sperm for fertilization. The role of autophagy in this process remains unclear. Here we provide evidence that pollen germination is accompanied by elevated autophagic activity and successful pollen tube emergence depends on autophagy-mediated cytoplasmic deletion. Genetic and cytological experiments demonstrate that inhibition of autophagy prevents pollen germination while induces the persistence of a layer of undegraded cytoplasm at the germination aperture. Together, these results unveil a novel compartmentalized autophagy. Furthermore, high-throughput comparative lipidomic analyses show that suppressed autophagy-induced inhibition of pollen germination is accompanied by altered profiles of stored and signaling lipids. Proteomic analyses reveal that autophagy likely exert its role in pollen germination via downstream mitochondria-related pathways. These findings reveal a critical role for autophagy in initiating pollen germination and provide evidences for compartmental cytoplasmic deletion being crucial for male fertility. Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy-related gene; Cer: ceramide; CL: cardiolipin; Con A: concanamycin A; DAG: diradylglycerol; GO: gene ontology; HAG: hour after germination; LC-MS: liquid chromatography-mass spectrometry; MAG: min after germination; MDC: monodansylcadaverine; PE: phosphatidylethanolamine; PI: phosphatidylinositol; PLD: phospholipase D; PtdIns3K: phosphatidylinositol 3-kinase; RT-qPCR: quantitative real-time reverse transcription PCR; TAG: triradylglycerol; TEM: transmission electron microscopy; TMT: tandem mass tagging.

[Applicability of Bioavailable Phosphorus in Sediments to Indicating Trophic Levels of Lakes and Reservoirs].

Twelve lakes and reservoirs with different water depths and different water residence times were studied to identify the applicability of bioavailable phosphorus of sediments in indicating trophic levels. Water and sediment samples were collected in these 12 lakes and reservoirs to analyze the relationship of nutrient levels between the sediment and the water column. Sodium hydroxide extracted phosphorus (NaOH-P) determined using the SMT classification method is defined as the bioavailable phosphorus of sediment. The results showed that total phosphorus levels in sediments in different lakes and reservoirs ranged from 225 to 760 mg·kg-1 (mean value 502 mg·kg-1); the NaOH-P levels in sediments ranged from 86 to 584 mg·kg-1 (mean value 263 mg·kg-1); the total phosphorus concentrations in the water was 0.02-0.35 mg·L-1 (mean value 0.11 mg·L-1), and the chlorophyll a concentrations in the water were 3-349 µg·L-1 (mean value 51 µg·L-1). It was found that NaOH-P was more effective than total phosphorus in indicating the trophic status of the lakes and reservoirs. However, the NaOH-P levels were significantly related to the phosphorus concentrations in the water column only in shallow water with a long residence time. It was revealed that water residence time and water depth are two key factors that affect the relationship of the phosphorus content between the sediment and the water column. In deep waters or waters with short residence time, the NaOH-P content in the sediment hardly influenced the phosphorus concentration in the water columns, even at high levels. However, in shallow waters with long residence time, the sediment acted as both sources and sinks and frequently exchanged nutrients with the overlying water, especially during bloom periods in summer. Thus NaOH-P could be a potential risk of eutrophication in such waters.

Efficacy and Safety of a Formulated Herbal Granula, Jiu Wei Zhen Xin, for Generalized Anxiety Disorder: A Meta-Analysis.

BACKGROUND: The traditional Chinese medicine formula Jiu Wei Zhen Xin Granula (JWZXG) is prescribed to treat generalized anxiety disorder (GAD) in China. This study was to assess the efficacy and safety of JWZXG in patients with GAD. METHOD: Data were pooled from 14 randomized controlled trials involving the assessment of mean changes of Hamilton Anxiety Rating Scale (HAMA) total scores, response rates, adverse event rates, quality, publication bias, and risk of bias. RESULTS: Pooled analysis showed no significant difference in response rate (risk ratio 1.01, 95% CI [0.93-1.08]; Z test = 0.17, P = 0.86) and no significant difference between JWZXG group and azapirones group (RR 0.69, 95% CI [0.45, 1.06]; Z test = 1.69, P = 0.09) in rate of adverse events. Though no difference exists between JWZXG group and azapirones group in HAMA total score from baseline, JWZXG group was inferior to selective serotonin reuptake inhibitors (SSRIs) group (WMD -0.93, 95% CI [-1.64, -0.23]; Z test = 2.6, P = 0.009) which had more adverse events than JWZXG group (RR 0.64, 95% CI [0.46, 0.89]; Z test = 2.63, P = 0.009). CONCLUSIONS: This meta-analysis preliminarily suggests that JWZXG is as effective as azapirones, though having the same possibility of suffering AEs. JWZXG was inferior to SSRIs but causes fewer AEs in the treatment of GAD.

Use of an UHPLC-MS/MS Method for Determination of Kuraridin and Characterization of Its Metabolites in Rat Plasma after Oral Administration.

Kuraridin is an active natural prenylated flavonoid ingredient originating from the well-known traditional Chinese medicine Sophora flavescens Ait., that possesses various bioactivities, such as antitumor activity, PLCγ1 inhibitory activity, glycosidase inhibitory activity, etc. However, there is no report on the plasma metabolic profile and pharmacokinetic study of kuraridin. The current study was designed to use an ultra-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) method for the quantification and characterization metabolites in rat plasma after oral administration of kuraridin. A liquid-liquid extraction method with ethyl acetate-acetonitrile (1:3) was used to extract the kuraridin from rat plasma samples. The chromatographic separation was carried out on a Hypersil GOLD UHPLC C18 column equipped with a C18 guard cartridge using a gradient elution with organic solvent-water as mobile phase. Based on comparing the retention times with reference standards or on the basis of MS2 fragmentation behaviors, a total of 19 metabolites were identified or tentatively characterized from rat plasma. Under the optimized conditions, the method showed good linearity (r² > 0.99) over the ranges of 1-500 ng/mL for kuraridin. The inter- and intra-day precisions were less than 8.95%, and the accuracy was in the range of -6.27-6.48%. The recovery of kuraridin ranged from 90.1% to 100.4%. The developed UHPLC-MS/MS method was thus successfully applied in the qualitative of metabolites and quantitative analysis of kuraridin in rat plasma.

[Virtual screening of anti-angiogenesis flavonoids from Sophora flavescens].

Angiogenesis is a dynamic, multi-step process. It is known that about 70 diseases are related to angiogenesis. Both the experimental and the literature reports showed that Sophora flavescens inhibit angiogenesis significantly, but the material basis and the mechanism of action have not been clear. In this study, molecular docking was used for screening of anti-angiogenesis flavonoids from the roots of S. flavescens. One handred and twenty-six flavonoids selected from S. flavescens were screened in the docking ligand database with six targets(VEGF-a,TEK,KDR,Flt1,FGFR1 and FGFR2) as the receptors. In addition, the small-molecule approved drugs of targets from DrugBank database were set as a reference with minimum score of each target's approved drugs as threshold. The LibDock module in Discovery Studio 2.5 (DS2.5) software was applied to screen the compounds. As a result, 37 compounds were screened out that their scores were higher than the minimum score of approved drugs as well as being in the top of 10%. At last the mechanism of flavonoids anti-angiogenesis was preliminarily revealed, which provided a new method for the development of angiogenesis inhibitor drugs.

[Water quality comparison between Lake Taihu and contribute river during high water-level period].

Thirty-two main rivers around Lake Taihu were sampled during high water period in 2008 to investigate river's vital impact on Lake water quality. Different forms of nitrogen and phosphorus were analyzed for the 32 river water samples according to nutrients function to algae. The aim of this study is to provide the basic information for river rehabilitation. Based on the results, the nutrients and suspended substance (SS) concentrations of Wangyu River were the highest, its water quality was below the V grade (Chinese water quality criterion). Water quality of northern rivers was also below the V grade, organic matter and enzymatically hydrolysable phosphate (EHP) concentration were the highest. The EHP concentration was 0.240 mg/L, and accounted for 62.4% of total phosphorus (TP). Nitrite nitrogen (NO2(-) -N) concentration was exceptionally high in western rivers and the average concentration was 0.270 mg/L, it was 2-3 times higher than that of other rivers. The northern rivers' nitrogen, phosphorus and organic concentration were markedly higher than those in the north Lake region, while the western rivers' nitrogen was markedly higher than that in the west Lake region. It can also be concluded that algae and suspended sediment were able to bring more phosphorus to water by comparing the ratio of total nitrogen(TN) and TP in these rivers and Lake Taihu.

[Study on effect of demethoxycurcumin in Curcuma long on stability of curcumin].

OBJECTIVE: To investigate effect of demethoxycurcumin on stability of curcumin. METHODS: To add the demethoxycurcumin to pure curcumin, the change of curcumin was determined by HPLC and the dynamics of curcumin degradation was investigated. RESULT: The stability both obtained from alcohol and demethoxycurcumin improved the stabilization of curcumin, the demi-period of curcumin prolonged with the addition of demethoxycurcumin. CONCLUSION: The commixture of curcumin and demethoxycurcumin are more stable than pure curcumin at the same conditions. Stability of curcumin is improved by demethoxycurcumin,it is crude stabilizing agent.

[Study on stability of curcumine, demethoxycurcumin and bisdemethoxycurcumin].

OBJECTIVE: To investigate the stability of curcumin, demethoxycurcumin and bisdemethoxycurcumin in different buffer solution. METHOD: To determine concentration of curcumin by HPLC when added curcumin, demethoxycurcumin and bisdemethoxycurcumin into the buffer solution the equation of degradation was established. RESULT: The sequence of stability are bisdemethoxycurcumin > or = demethoxycurcumin > or =curcumin at the same condition. CONCLUSION: The demethoxycurcumin can stabilize curcumin more strong than the others. The demethoxycurcumin is a nature stabilizing agent for curcumin.