Unlocking the potential of amorphous calcium carbonate: A star ascending in the realm of biomedical application.

Calcium-based biomaterials have been intensively studied in the field of drug delivery owing to their excellent biocompatibility and biodegradability. Calcium-based materials can also deliver contrast agents, which can enhance real-time imaging and exert a Ca2+-interfering therapeutic effect. Based on these characteristics, amorphous calcium carbonate (ACC), as a brunch of calcium-based biomaterials, has the potential to become a widely used biomaterial. Highly functional ACC can be either discovered in natural organisms or obtained by chemical synthesis However, the standalone presence of ACC is unstable in vivo. Additives are required to be used as stabilizers or core-shell structures formed by permeable layers or lipids with modified molecules constructed to maintain the stability of ACC until the ACC carrier reaches its destination. ACC has high chemical instability and can produce biocompatible products when exposed to an acidic condition in vivo, such as Ca2+ with an immune-regulating ability and CO2 with an imaging-enhancing ability. Owing to these characteristics, ACC has been studied for self-sacrificing templates of carrier construction, targeted delivery of oncology drugs, immunomodulation, tumor imaging, tissue engineering, and calcium supplementation. Emphasis in this paper has been placed on the origin, structural features, and multiple applications of ACC. Meanwhile, ACC faces many challenges in clinical translation, and long-term basic research is required to overcome these challenges. We hope that this study will contribute to future innovative research on ACC.

The adsorbent preparation of lanthanum functionalized sponge based on CMC coating for effective phosphorous removal.

Eutrophication is a severe worldwide concern caused by excessive phosphorus release. Thus, significant efforts have been made to develop phosphorus removal techniques, particularly by nanomaterial adsorption. However, because of the limitations associated with nanoparticles including easy agglomeration, and separation challenges, a novel nanocomposite adsorbent with great adsorption performance is urgently required. A sponge adsorbent (MS-CMC@La) was developed in this study to remove phosphorus using melamine sponge (MS), LaCl3, and sodium carboxymethyl cellulose (CMC). The results of SEM/EDS, FTIR, and XPS demonstrated that La was well-dispersed on MS-CMC@La. Adsorption isotherm and kinetics met with the Langmuir model (R2 = 0.981) and the pseudo-second-order kinetics (R2 = 0.989), respectively. The maximum adsorption capacity of MS-CMC@La was found to be 15.28 mg/g; the material exhibited excellent selectivity toward phosphorus in the presence of coexisting anion except of F-; the adsorption behavior was greatly impacted by pH. Furthermore, the electrostatic attraction, ligand exchange and inner-sphere coordination regulate the phosphate adsorption mechanism, with inner-sphere coordination dominating. In summary, the nano-enriched materials developed in this study are capable of facilitating the application of functionalized sponges in the field of wastewater.

Isoandrographolide from Andrographis paniculata ameliorates tubulointerstitial fibrosis in ureteral obstruction-induced mice, associated with negatively regulating AKT/GSK-3ß/ß-cat signaling pathway.

Tubulointerstitial fibrosis (TIF) is a prominent pathological manifestation for the progression of almost all chronic kidney diseases (CKDs) to end-stage renal failure. However, there exist few efficient therapies to cure TIF. Our recent results showed that (8R, 12S)-isoandrographolide (ISA), a diterpenoid lactone ingredient of traditional Chinese herbal Andrographis paniculata (Burm.f.) Nees, exhibited anti-pulmonary fibrosis in silica-induced mice. Herein, we investigated the therapeutic effect of ISA on TIF, using mice subjected to unilateral ureteral obstruction (UUO) and human kidney proximal tubular epithelial (HK-2) cells treated with transforming growth factor-ß1 (TGF-ß1) or tumor necrosis factor-α (TNF-α). The pathological changes and collagen deposition results displayed that ISA administration significantly attenuated inflammatory response, ameliorated TIF, and protected the kidney injury. Interestingly, ISA revealed much lower cytotoxicity on HK-2 cells, but exhibited stronger inhibitory effect on tubular epithelial mesenchymal transformation (EMT) and inflammation, as compared to andrographolide (AD), the major ingredient of A. paniculata extract that has been reported to ameliorate TIF in diabetic nephropathy mice. It was further clarified that the amelioration of TIF by ISA was associated with suppressing the aberrant activation of AKT/GSK-3ß/ß-catenin pathway through network pharmacology analysis and experimental validation. Taken together, these findings indicate that ISA is a promising lead compound for development of anti-TIF, and even broad-spectrum anti-fibrotic drugs.

Genome Wide Identification and Characterization of Wheat GH9 Genes Reveals Their Roles in Pollen Development and Anther Dehiscence.

Glycoside hydrolase family 9 (GH9) is a key member of the hydrolase family in the process of cellulose synthesis and hydrolysis, playing important roles in plant growth and development. In this study, we investigated the phenotypic characteristics and gene expression involved in pollen fertility conversion and anther dehiscence from a genomewide level. In total, 74 wheat GH9 genes (TaGH9s) were identified, which were classified into Class A, Class B and Class C and unevenly distributed on chromosomes. We also investigated the gene duplication and reveled that fragments and tandem repeats contributed to the amplification of TaGH9s. TaGH9s had abundant hormone-responsive elements and light-responsive elements, involving JA-ABA crosstalk to regulate anther development. Ten TaGH9s, which highly expressed stamen tissue, were selected to further validate their function in pollen fertility conversion and anther dehiscence. Based on the cell phenotype and the results of the scanning electron microscope at the anther dehiscence period, we found that seven TaGH9s may target miRNAs, including some known miRNAs (miR164 and miR398), regulate the level of cellulose by light and phytohormone and play important roles in pollen fertility and anther dehiscence. Finally, we proposed a hypothesis model to reveal the regulation pathway of TaGH9 on fertility conversion and anther dehiscence. Our study provides valuable insights into the GH9 family in explaining the male sterility mechanism of the wheat photo-thermo-sensitive genetic male sterile (PTGMS) line and generates useful male sterile resources for improving wheat hybrid breeding.

Isoandrographolide inhibits NLRP3 inflammasome activation and attenuates silicosis in mice.

Silicosis is an irreversible occupational disease caused by silica particle exposure. Abundant evidences suggest that NLRP3-mediated inflammation acts an essential role in fibrogenesis and the pathogenesis of silicosis. In the current work, we firstly reported that (8R-12S)-isoandrographolide (ISA), a diterpenoid lactone ingredient of Chinese traditional medicinal plant Andrographis paniculata (Burm.f.) Nees, could reduce pulmonary inflammation and fibrosis by inhibiting NLRP3, and thereby ameliorate silicosis. ISA administration significantly alleviated lung injury, and attenuated inflammatory response, EMT, as well as collagen deposition in the lung of silica-induced mice. Further studies verified that ISA inhibited the expressions of NLRP3 inflammasome-related proteins NLRP3, ASC and caspase-1 in vivo and in vitro, leading to the attenuation of inflammation and EMT. Additionally, the molecular docking assay indicated that ISA possibly interacted with the residues of LYS26 and GLU47 of NLRP3, implying that ISA might directly bond to protein NLRP3. Of note, ISA revealed a lower cytotoxicity but more potent therapeutic effect than andrographolide (AD), the major active extract of A. paniculata, which has been traditionally used to treat inflammation-related diseases. Taken together, our study clarified a novel role of ISA in attenuating inflammation and fibrosis in silicosis, and indicated a bright future of ISA as a lead compound for developing therapeutic drug for silicosis.

Comprehensive analysis of formin gene family highlights candidate genes related to pollen cytoskeleton and male fertility in wheat (Triticum aestivum L.).

BACKGROUND: Formin, a highly conserved multi-domain protein, interacts with microfilaments and microtubules. Although specifically expressed formin genes in anthers are potentially significant in research on male sterility and hybrid wheat breeding, similar reports in wheat, especially in thermo-sensitive genic male sterile (TGMS) wheat, remain elusive. RESULTS: Herein, we systematically characterized the formin genes in TGMS wheat line BS366 named TaFormins (TaFHs) and predicted their functions in inducing stress response. In total, 25 TaFH genes were uncovered, majorly localized in 2A, 2B, and 2D chromosomes. According to the neighbor-joining (NJ) method, all TaFH proteins from wheat and other plants clustered in 6 sub-groups (A-F). The modeled 3D structures of TaFH1-A/B, TaFH2-A/B, TaFH3-A/B and TaFH3-B/D were validated. And different numbers of stress and hormone-responsive regulatory elements in their 1500 base pair promoter regions were contained in the TaFH genes copies. TaFHs had specific temporal and spatial expression characteristics, whereby TaFH1, TaFH4, and TaFH5 were expressed highly in the stamen of BS366. Besides, the accumulation of TaFHs was remarkably lower in a low-temperature sterile condition (Nanyang) than fertile condition (Beijing), particularly at the early stamen development stage. The pollen cytoskeleton of BS366 was abnormal in the three stages under sterile and fertile environments. Furthermore, under different stress levels, TaFHs expression could be induced by drought, salt, abscisic acid (ABA), salicylic acid (SA), methyl jasmonate (MeJA), indole-3-acetic acid (IAA), polyethylene glycol (PEG), and low temperature. Some miRNAs, including miR167, miR1120, and miR172, interacts with TaFH genes; thus, we constructed an interaction network between microRNAs, TaFHs, phytohormone responses, and distribution of cytoskeleton to reveal the regulatory association between upstream genes of TaFH family members and sterile. CONCLUSIONS: Collectively, this comprehensive analysis provides novel insights into TaFHs and miRNA resources for wheat breeding. These findings are, therefore, valuable in understanding the mechanism of TGMS fertility conversion in wheat.

[Research progress of Astragali Radix fermentation].

Astragali Radix is one of traditional Chinese medicines with effects in invigorating Qi for consolidating superficies, inducing diuresis to alleviate edema, promoting pus discharge and tissue regeneration. In recent years, the traditional Chinese medicine fermentation technology has received extensive attentions due to its high efficiency and safety. The pharmacological functions of traditional Chinese medicines could be further enhanced after microbial fermentation, which has a broad development prospects. In this paper, we summarized relevant literatures of Astragali Radix fermentation in such aspects as fermentation strains, fermentation forms, process optimization, active ingredients and pharmacological effects, in the expectation of providing a reference for development and utilization of Astragali Radix.

A stepwise integrated multi-system to screen quality markers of Chinese classic prescription Qingzao Jiufei decoction on the treatment of acute lung injury by combining 'network pharmacology-metabolomics-PK/PD modeling'.

BACKGROUND: Previously, we have investigated the therapeutic mechanism of Qingzao Jiufei Decoction (QZJFD), a Chinese classic prescription, on acute lung injury (ALI), however, which remained to be further clarified together with the underlying efficacy related compounds for quality markers (Q-markers). HYPOTHESIS/PURPOSE: To explore Q-markers of QZJFD on ALI by integrating a stepwise multi-system with 'network pharmacology-metabolomics- pharmacokinetic (PK)/ pharmacodynamic (PD) modeling'. METHODS: First, based on in vitro and in vivo component analysis, a network pharmacology strategy was developed to identify active components and potential action mechanism of QZJFD on ALI. Next, studies of poly-pharmacology and non-targeted metabolomics were used to elaborate efficacy and verify network pharmacology results. Then, a comparative PK study on active components in network pharmacology was developed to profile their dynamic laws in vivo under ALI, suggesting Q-marker candidates. Next, quantified analytes with marked PK variations after modeling were fitted with characteristic endogenous metabolites along drug concentration-efficacy-time curve in a PK-PD modeling to verify and select primary effective compounds. Finally, Q-markers were further chosen based on representativeness among analytes through validity analysis of PK quantitation of primary effective compounds. RESULTS: In virtue of 121 and 33 compounds identified in vitro and in vivo, respectively, 33 absorbed prototype compounds were selected to construct a ternary network of '20 components-47 targets-113 pathways' related to anti-ALI of QZJFD. Predicted mechanism (leukocytes infiltration, cytokines, endogenous metabolism) were successively verified by poly-pharmacology and metabolomics. Next, 18 measurable components were retained from 20 analytes by PK comparison under ALI. Then, 15 primary effective compounds from 18 PK markers were further selected by PK-PD analysis. Finally, 9 representative Q-markers from 15 primary effective compounds attributed to principal (chlorogenic acid), ministerial (methylophiopogonanone A, methylophiopogonanone B), adjuvant (sesamin, ursolic acid, amygdalin), conductant drugs (liquiritin apioside, liquiritigenin and isoliquiritin) in QZJFD, were recognized by substitutability and relevance of plasmatic concentration at various time points. CONCLUSION: 9 Q-markers for QZJFD on ALI were identified by a stepwise integration strategy, moreover, which was a powerful tool for screening Q-makers involved with the therapeutic action of traditional Chinese medicine (TCM) prescription and promoting the process of TCM modernization and scientification.

Acute lung injury therapeutic mechanism exploration for Chinese classic prescription Qingzao Jiufei Decoction by UFLC-MS/MS quantification of bile acids, fatty acids and eicosanoids in rats.

Acute lung injury (ALI) is a common and complex inflammatory disease, which has been reasonably associated with carboxyl-containing metabolites in our preliminary non-targeted metabolomic strategy. Qingzao Jiufei Decoction (QZJFD), a classic prescription, is widely used in the treatment of pulmonary inflammatory injuries. Successively, in this targeted project, to fill in the research gap and exposit the therapeutic mechanism of QZJFD on ALI, considering the structure similarity and bioactivity correlation, 21 bile acids, 11 fatty acids and 19 eicosanoids were profiled simultaneously in plasma, lung, bronchoalveolar lavage fluid, spleen and feces from rats utilizing a novel ultraperformance liquid chromatography-mass spectrometry approach. As a result, potential biomarkers and ALI characteristic metabolomic spectrums were obtained to distinguish different physical states using discriminative similarity threshold as 0.65 for clinical application. After treatment with QZJFD, obvious reversing ability for various biomarker levels was observed in different bio-samples, providing insights into the systemic intervention of QZJFD on ALI by regulating bile acid synthesis, fatty acid synthesis and eicosanoid metabolism. Conclusively, this investigation represented more information on the comprehensive therapeutic action of QZJFD on ALI involving with multi-targets and multi-pathways for clinical application and traditional Chinese medicine modernization.

The chemical transformations for Radix Astragali via different alkaline wash conditions by quantitative and qualitative analyses.

Radix Astragali is a famous Chinese traditional and folk medicine with a wide range of medicinal values in clinic. In this study, an analytical efficient strategy based on UHPLC-QQQ-MS/MS and UHPLC-LTQ-Orbitrap-MS/MS was established to explore and reveal the chemical transformations for Radix Astragali under different alkaline wash conditions for analytical sample preparation. Firstly, a rapid and sensitive UHPLC-QQQ-MS/MS method for the quantification of 14 main constituents in Radix Astragali has been developed and validated. Secondly, according to the standard substance comparison, accurate mass measurements, mass fragmentation behaviors and related literatures, a total of 102 components have been screened and identified using UHPLC-LTQ-Orbitrap method. Among them, 47 compounds are saponins, and the other 55 are flavonoids. Consequently, there were two chemical transformations including hydrolysis and degradation observed when Radix Astragali was treated with alkali. Besides, hydrolysis of glycosides and acetyl played a considerably important role in the process of sample preparation. It has been proved that 10 % ammonia could relatively guarantee the high content of astragaloside IV and avoid the over-degradation of most chemical ingredients in Radix Astragali. In conclusion, this work would provide a scientific and practical method for quality control of Radix Astragali as well as its compound preparations.

Comprehensive analysis of the chemical constituents in sulfur-fumigated Lonicerae Japonicae Flos using UHPLC-LTQ-Orbitrap mass spectrometry.

Lonicerae Japonicae Flos (LJF), a kind of traditional Chinese medicines (TCMs), has functions of detoxifying and evacuating heat. In the study, a method based on ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap MS) was developed for the chemical constituent analysis of organic acids, flavonoids, iridoids and new-generated compounds in sulfur-fumigated LJF (SF-LJF). Based on the accurate mass measurement (< ± 5 ppm), chromatographic behavior and diagnostic product ions (DPIs), 113 constituents were unambiguously or tentatively characterized from SF-LJF extract, including 46 chlorogenic acids, 19 flavonoids, 29 iridoid glycosides and 19 newly-generated compounds (including 17 sulfur-containing derivatives). In addition, 5-CQA (5-caffeoylquinic acid, chlorogenic acid) was chosen to be sulfur-fumigated for the result validation. It was found that the most significant change of LJF after sulfur fumigation was the occurrence of sulfate or sulfite esterification reactions, which resulted in the emergence of many new sulfur-containing components. Our results demonstrated that the established method was a useful and efficient analytical tool to comprehensively characterize the material basis of SF-LJF, and also an excellent guidance of quality control about LJF.

Comparative transcriptome analysis indicates conversion of stamens into pistil-like structures in male sterile wheat (Triticum aestivum L.) with Aegilops crassa cytoplasm.

BACKGROUND: Aegilops crassa cytoplasm is an important source for investigating cytoplasmic male sterility (CMS). Moreover, the stamens of line C303A exhibit a high degree of pistillody, turning almost white. However, the molecular mechanism that underlies pistillody in C303A remains unclear. Therefore, to obtain a better understanding of pistillody in C303A, the phenotypic and cytological features of C303A were observed to identify the key stage for the homeotic transformation of stamens into pistil-like structures. Transcriptome profiles were determined for stamens using Illumina RNA sequencing. RESULTS: Morphological observations of the CMS wheat line with Aegilops crassa cytoplasm C303A showed that the pistils developed normally, but the stamens were ultimately aborted and they released no pollen when mature. According to paraffin section observations, the stamens began to transform into pistils or pistil-like structures in the binucleate stage (BNS). Therefore, the stamens were collected from line C303A and its maintainer 303B in the BNS for transcriptome sequencing. In total, 20,444 wheat genes were determined as differentially expressed in C303A and 303B stamens, with 10,283 upregulated and 10,161 downregulated genes. Gene Ontology enrichment analyses showed that most of the differentially expressed genes (DEGs) were annotated with GO terms comprising metabolic process, cell, cellular process, catalytic activity, and cell part. Analysis based on the Kyoto Encyclopedia of Genes and Genomes database showed that the enriched DEGs were mainly associated with energy metabolism. We also found several essential genes that may contribute to pistillody in C303A. These findings suggest that disrupted energy metabolism and reactive oxygen metabolism induce pistillody and eventually lead to abortion in C303A. CONCLUSION: We determined the complex transcriptome profiles for C303A stamens and demonstrated that disrupted energy metabolism and class B MADS-box genes are related to pistillody. These findings may facilitate future studies of the mechanistic response of the wheat stamen and pollen development in CMS.

Metabolomic profile perturbations of serum, lung, bronchoalveolar lavage fluid, spleen and feces in LPS-induced acute lung injury rats based on HPLC-ESI-QTOF-MS.

Acute lung injury (ALI) is a clinically common and serious disease, underscoring the urgent need for clarification of its pathogenesis. According to traditional Chinese medicine (TCM) theories on the "lung-spleen-intestine axis" and its correlation with ALI, a high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) metabolomic platform was applied to identify biomarkers from five bio-samples of control and model rats challenged with intratracheally administered lipopolysaccharide (LPS) based on multivariate mathematical statistical analysis. As a result, 19, 24, 24, 15 and 29 altered metabolites were identified in serum, lung, bronchoalveolar lavage fluid (BALF), spleen and feces samples, respectively. Metabolic pathway analysis showed that linoleic acid, sphingolipid, glycerophospholipid and bile acid metabolism pathways were mainly altered by ALI. Additionally, ROC curves were applied to assess the specificity and sensitivity of the biomarkers. ALI characteristic metabolomic spectra were then established to differentiate the control from the model group with a similarity discriminative threshold of 0.7. Additionally, to compare the metabolomic profiles of the five bio-samples and establish metabolic similarities and differences among them, correlation analysis was conducted in order to delineate an objective law of endogenous linkage along the lung-spleen-intestine axis. Therefore, this study provides insights into the mechanisms involved in ALI from a metabolomics perspective, which can be applied in characterization of the mechanism and early disease detection. Graphical abstract.

Comparative transcriptome analysis indicates that a core transcriptional network mediates isonuclear alloplasmic male sterility in wheat (Triticum aestivum L.).

BACKGROUND: Cytoplasmic male sterility (CMS) plays a crucial role in the utilization of heterosis and various types of CMS often have different abortion mechanisms. Therefore, it is important to understand the molecular mechanisms related to anther abortion in wheat, which remain unclear at present. RESULTS: In this study, five isonuclear alloplasmic male sterile lines (IAMSLs) and their maintainer were investigated. Cytological analysis indicated that the abortion type was identical in IAMSLs, typical and stainable abortion, and the key abortive period was in the binucleate stage. Most of the 1,281 core shared differentially expressed genes identified by transcriptome sequencing compared with the maintainer in the vital abortive stage were involved in the metabolism of sugars, oxidative phosphorylation, phenylpropane biosynthesis, and phosphatidylinositol signaling, and they were downregulated in the IAMSLs. Key candidate genes encoding chalcone--flavonone isomerase, pectinesterase, and UDP-glucose pyrophosphorylase were screened and identified. Moreover, further verification elucidated that due to the impact of downregulated genes in these pathways, the male sterile anthers were deficient in sugar and energy, with excessive accumulations of ROS, blocked sporopollenin synthesis, and abnormal tapetum degradation. CONCLUSIONS: Through comparative transcriptome analysis, an intriguing core transcriptome-mediated male-sterility network was proposed and constructed for wheat and inferred that the downregulation of genes in important pathways may ultimately stunt the formation of the pollen outer wall in IAMSLs. These findings provide insights for predicting the functions of the candidate genes, and the comprehensive analysis of our results was helpful for studying the abortive interaction mechanism in CMS wheat.

[UHPLC-HRMS metabonomics to study effect of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats].

The project was launched to analyze the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats by metabonomics. The preparation method of sulfur-fumigated Ophiopogonis Radix in laboratory was established. Then the blood samples of SD rats in blank group,Ophiopogonis Radix extract group and sulfur-fumigated Ophiopogonis Radix extract group were investigated by UHPLC-Q-Exactive. The differential metabolites were screened and identified by PCA(principal component analysis),OPLSDA(orthogonal partial least squares discriminant analysis) and variable importance projection(VIP),and the metabolic pathways were analyzed. Finally,a total of 15 potential biomarkers were identified. Compared with the samples of Ophiopogonis Radix extract group,sulfur-fumigated Ophiopogonis Radix mainly affected the biosynthesis and metabolism of amino acids in normal rats. Its mechanism may be related to the biosynthesis of phenylalanine,tyrosine,tryptophan and aminoacyl-tRNA as well as the metabolism of phenylalanine and tryptophan. Based on UHPLC-HRMS metabonomics,this paper discussed the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats,which provided an idea for the metabolic study of other sulfur-fumigated traditional Chinese medicines.

Chemical Constituent Profiling of Paecilomyces cicadae Liquid Fermentation for Astragli Radix.

Astragli Radix (AR) is one of the most popular traditional Chinese medicines with chemical constituents including flavonoids and saponins. As recently evidenced, some fungi or their fermentation liquid may have the potential to affect the bioactive constituents and different pharmacological effects of AR. Thus, the composition of fermented AR (FAR) produced by Paecilomyces cicadae (Miquel) Samson in liquid-state fermentation was investigated using a UHPLC-LTQ-Orbitrap mass spectrometer in both positive and negative ion modes. Firstly, the MSn data sets were obtained based on a data-dependent acquisition method and a full scan-parent ions list-dynamic exclusion (FS-PIL-DE) strategy. Then, diagnostic product ions (DPIs) and neutral loss fragments (NLFs) were proposed for better constituent detection and structural characterization. Consequently, 107 constituents in total, particularly microconstituents in FAR and AR, were characterized and compared in parallel on the same LTQ-Orbitrap instrument. Our results indicated that AR fermentation with Paecilomyces significantly influenced the production of saponins and flavonoids, especially increasing the content of astragaloside IV. In conclusion, this research was not only the first to show changes in the chemical components of unfermented AR and FAR, but it also provides a foundation for further studies on the chemical interaction between microbiota and AR.

Temperature-dependent growth and hypericin biosynthesis in Hypericum perforatum.

Hypericum perforatum is a perennial herb that produces the anti-depression metabolite hypericin (Hyp). While several efforts to increase Hyp production have been made, the effects of temperatures on growth and Hyp biosynthesis are still limited. In this study, the growth morphophysiological traits, Hyp biosynthesis and their related genes expression, as well as major bioactive compounds accumulation and antioxidant capacity were assessed by exposing H. perforatum seedlings to three different temperatures (15, 22 and 30 °C). The results showed that aerial parts biomass was greater at 15 °C with 1.3 and 1.6-fold increase compared to at 22 and 30 °C, in large part because of greater increase in chlorophyll content, stem number and leaf area on a per plant basis. Hyp content in the aerial parts was greater 1.9 and 5.6-fold on a per plant basis compared to 22 and 30 °C treatments, and the contents of other bioactive compounds (flavonoids and phenolics) as well as antioxidant capacity in the aerial parts, on dry weight and per plant basis, also exhibited significant increases with the temperatures decrease. The mRNA expressions of eight genes (psbA, psbB, psbC, psbD, ycf3, ycf4, ycf5 and matK) related to photosynthesis and two genes (Polyketide synthase, PKS; Phenolic oxidative coupling protein, Hyp-1) involved in Hyp biosynthesis were also up-regulated at 15 °C. The findings are useful in guiding cultivation and regulating Hyp biosynthesis in H. perforatum.

The complete chloroplast genome of Tagetes erecta (Asteroideae), a wildly cultivated ornamental plant.

Tagetes erecta (Asteraceae) has been wildly cultivated as ornamental and medicinal plant. Here, we reported the first chloroplast genome sequence of T. erecta. The chloroplast genome size is 152,065 bp with GC content of 37.4%, including a large single-copy (LSC) of 83,895 bp, a small single-copy (SSC) of 18,065 bp, and a pair of 25,048 bp IR (inverted repeat) regions. A total of 132 genes were annotated including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic analysis revealed that T. erecta belongs to the subfamily Asteroideae.

Tapetal-Delayed Programmed Cell Death (PCD) and Oxidative Stress-Induced Male Sterility of Aegilops uniaristata Cytoplasm in Wheat.

Cytoplasmic male sterility (CMS) plays a crucial role in the utilization of hybrid vigor. Pollen development is often accompanied by oxidative metabolism responses and tapetal programmed cell death (PCD), and deficiency in these processes could lead to male sterility. Aegilops uniaristata cytoplasmic male sterility (Mu-CMS) wheat is a novel male-sterile line in wheat, which possess important potential in hybrid wheat breeding. However, its CMS mechanisms remain poorly understood. In our study, U87B1-706A, with the Aegilops uniaristata cytoplasm, and the maintainer line 706B were used to explore the abortive reason. Compared with 706B, histological analysis and PCD detection of the anther demonstrated that U87B1-706A appeared as delayed tapetal PCD as well as a disorganized organelle phenotype in the early uninucleate stage. Subsequently, a shrunken microspore and disordered exine structure were exhibited in the late uninucleate stage. While the activities of antioxidase increased markedly, the nonenzymatic antioxidant contents declined obviously following overacummulation of reactive oxygen species (ROS) during pollen development in U87B1-706A. Real-time quantitative PCR testified that the transcript levels of the superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) genes, encoding pivotal antioxidant enzymes, were up-regulated in early pollen development. Therefore, we deduce excess ROS as a signal may be related to the increased expression levels of enzyme genes, thereby breaking the antioxidative system balance, resulting in delayed tapetal PCD initiation, which finally led to pollen abortion and male sterility in U87B1-706A. These results provide evidence to further explore the mechanisms of abortive pollen in CMS wheat.

[Qualitative and quantitative analysis of Evodiae Fructus based on the UPLC technology].

An UPLC method was developed for the studies of fingerprint and quantification of multi-components for Evodiae Fructus. The chromatographic separation was performed on a C18 column (2.1 mm×50 mm,1.7 µm) with mobile phase of 0.2% formic acid-acetonitrile and 0.2% formic acid-water in gradient mode, and the detection wavelength was set at 320 nm．Dehydroevodiamine was used as the reference peak, there were 24 common peaks in the fingerprint of 29 samples were detected, and among them 10 chromatographic peaks were identified with the reference substance and they were neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, hyperin, isorhamnetin-3-O-ß-D-rutinoside, dehydroevodiamine, evodiamine, rutaecarpine, evocarpine and dihydroevocarpine. The fingerprint data was evaluated with similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (Version 2008A), and the similarity of 19 batches of Evodiae Fructus was greater than 0.9 in the 29 samples. In addition, 9 components including neochlorogenic acid, chlorogenic acid, hyperin, isorhamnetin-3-O-ß-D-rutinoside, dehydroevodiamine, evodiamine, rutaecarpine, evocarpine and dihydroevocarpine were simultaneously determined at the same chromatographic conditions, whose peak area integral values showed good linear relationship at the range of 0.000 46-0.138, 0.000 146-0.175, 0.000 412-0.124, 0.000 448-0.134, 0.000 452-0.136, 0.003 38-0.169, 0.000 44-0.132, 0.001 07-0.128, 0.001 71-0.128, respectively. Their average recoveries were 100.3%, 100.4%, 101.6%, 97.51%, 102.9%, 101.4%, 103.8%, 104.0%, 95.99%, and RSD were 2.4%, 2.0%, 3.0%, 0.80%, 1.9%, 2.1%, 1.1%, 2.2%, 2.4%, respectively. The established UPLC method not only realized the full separation of all chemical constituents of Evodiae Fructus within 20 minutes, but also achieved the chromatographic fingerprint determination and simultaneous multi-components determination of Evodiae Fructus at the same chromatographic conditions. Compared with other methods in literatures, the method has the following characteristics of strong specificity, good separation, high purity of chromatographic peaks, simplity and feasibility, which provides better means for the simultaneous qualitative and quantitative analysis of Evodiae Fructus.