An integrated strategy using LC-MS/MS combined with in vivo microdialysis for the simultaneous determination of lignans of Schisandra chinensis (Turcz.) Baill. Fructus and endogenous neurotransmitters: application in pharmacokinetic and pharmacodynamic studies.

Schisandra chinensis (Turcz.) Baill Fructus (SCF) is the ripe fruit of Schisandra chinensis (Turcz.) Baill, and is often used as a neuroprotective drink. Modern pharmacological studies have shown that lignans are the main bioactive components responsible for neuroprotection and have potential in the treatment of Alzheimer's disease (AD). However, the mechanism of action of SCF in the treatment of AD from the pharmacokinetics-pharmacodynamics (PK-PD) perspective remains not well established. The purpose of this study is to investigate and compare the pharmacokinetic differences of lignans in normal and AD rats, as well as to investigate their effects on neurotransmitters and their role in the treatment of AD. To achieve this goal, an integrated strategy using LC-MS/MS combined with in vivo microdialysis for the simultaneous determination of lignans of SCF and endogenous neurotransmitters has been developed and validated. The results show that the pharmacokinetic behaviors of ten lignans in the AD group were significantly different from those in the normal group. The AD group had better absorption and slower elimination than the normal group. In addition, the pharmacodynamic results of the Morris water maze (MWM) test, biochemical tests, histopathological examination, as well as immunohistochemistry analysis showed that lignans could improve the learning and memory of AD rats. The oral administration of SCF could restore the levels of the neurotransmitter parameters; seven neurotransmitters showed clockwise or counterclockwise changes with the four lignans in the hippocampal region. Taken together, the PK and PD studies based on in vivo microdialysis sampling might offer novel insights into the mechanisms of action of SCF against AD.

Based on urine metabolomics to study the mechanism of Qi-deficiency affecting type 2 diabetes rats using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

Qi-deficiency also called energy deficiency, which approximates to the term of sub-health in contemporary medical theory. Diabetes is similar to the symptoms of "xiaoke" in traditional Chinese medicine (TCM) which is linked with Qi-deficiency. However, the mechanism of Qi-deficiency on type 2 diabetes (T2D) has not been completely elucidated. In this study, a model on Qi-deficiency T2D rat was established by using diet with high fat and high sugar and small-dose STZ induction combined with exhaustive swimming, and the model was evaluated by pathological section, hematological index and serum biochemical parameters. Applying urine metabolomics based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to explore the underlying molecular mechanism of Qi-deficiency on T2D and 32 urinary metabolites were identified as prospective biomarkers for Qi-deficiency T2D rats. Metabolic pathway analysis indicated that synthesis and degradation of ketone bodies, starch and sucrose metabolism, phenylalanine metabolism, arachidonic acid metabolism, butanoate metabolism and TCA cycle, etc., were closely related to potential mechanisms of Qi-deficiency on T2D. The metabolomics results can provide reliable data support for complex TCM syndrome diagnosis.

Mass spectrometry-based serum lipidomics strategy to explore the mechanism of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. leaves in the treatment of ischemic stroke.

Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. leaves (ESL) were reported to have neuroprotective function and are also used to treat cranial and cerebral traumas as a traditional Chinese medicine and food herbage plant. However, there has been no previous study on ESL treatment for stroke at the level of lipid disorders. To clarify the mechanism of ESL in treating ischemic stroke, this study was carried out from 3 aspects, namely, the regulation of lipid disorders, protection of the nervous system, as well as anti-inflammatory and antioxidant actions. This study established a lipidomics research strategy that was developed by UPLC-Q-TOF/MS analysis. The quantification of neurotransmitters in the serum and brain tissue of rats was performed using UPLC-TQ/MS. Also, we quantified the oxidative stress and inflammatory reaction by measuring the contents of SOD, MDA, TNF-α, IL-6, and IL-10 via the ELISA kits for serum and brain tissue. According to UPLC-Q-TOF/MS-based lipidomics analysis, 27 lipidomics biomarkers were identified in this study, including PC, PE, SM, and TG, which were distributed in various lipid metabolic pathways, including glycerophospholipid, linoleic acid, alpha-linolenic acid, glycerolipid, sphingolipid, and arachidonic acid metabolism pathways. By reversing the changes in the lipid content caused by the disease, ESL has a therapeutic effect on ischemic stroke. Furthermore, quantitative results of neurotransmitters indicated that they can be regulated by ESL. Finally, the results of ELISA showed that ESL can treat ischemic stroke to a certain extent by reducing the oxidative and inflammatory damage. Therefore, ESL may play a therapeutic role in the treatment of ischemic stroke in different ways. This research preliminarily revealed the mechanism of ESL in the treatment of ischemic stroke and provided support for the further application of ESL.

A rapid protocol to distinguish between Citri Exocarpium Rubrum and Citri Reticulatae Pericarpium based on the characteristic fingerprint and UHPLC-Q-TOF MS methods.

Citri Exocarpium Rubrum (CER) and Citri Reticulatae Pericarpium (CRP) are used as common functional foods and traditional Chinese medicine (TCM). As different parts of the same fruit, CER and CRP have different effects in clinical applications. However, they are commonly confused due to the similarity of the chemical compounds and a lack of scientific method to distinguish them in the finished product. In this study, an ultra-high performance liquid chromatography-diode array detector (UHPLC-DAD) technique and an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) method were employed to generate the characteristic fingerprint under the optimum analytical conditions. Principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were applied to represent different chemical markers for CER and CRP. 44 potential markers including 15 polymethoxylated flavanones (PMFs), 5 flavone-C-glycosides, 6 flavanone-O-glycosides, 3 flavonoid-O-glycosides, 8 organic acids, 5 limonoids and 2 alkaloids were successfully identified by using UNIFI software. The heat map showed that there were significant differences in the CER and CRP samples. Furthermore, 12 potential markers were screened out by characteristic fingerprint and UHPLC-Q-TOF MS methods and were analyzed by quantitative analysis of multicomponents by a single marker (QAMS) method. Finally, a prediction model based on the discovered chemical markers was established for discrimination between CER and CRP. Using these markers can significantly distinguish the unknown processed products of CER and CRP. In conclusion, an effective way to quickly and easily distinguish CER and CRP was successfully established based on the characteristic fingerprint and UHPLC-Q-TOF MS. It could also be a new strategy for analysis of different processed products of the same plant source.

Transcriptome Profiling of Wheat Inflorescence Development from Spikelet Initiation to Floral Patterning Identified Stage-Specific Regulatory Genes.

Early reproductive development in cereals is crucial for final grain number per spike and hence the yield potential of the crop. To date, however, no systematic analyses of gene expression profiles during this important process have been conducted for common wheat (Triticum aestivum). Here, we studied the transcriptome profiles at four stages of early wheat reproductive development, from spikelet initiation to floral organ differentiation. K-means clustering and stage-specific transcript identification detected dynamically expressed homeologs of important transcription regulators in spikelet and floral meristems that may be involved in spikelet initiation, floret meristem specification, and floral organ patterning, as inferred from their homologs in model plants. Small RNA transcriptome sequencing discovered key microRNAs that were differentially expressed during wheat inflorescence development alongside their target genes, suggesting that miRNA-mediated regulatory mechanisms for floral development may be conserved in cereals and Arabidopsis. Our analysis was further substantiated by the functional characterization of the ARGONAUTE1d (AGO1d) gene, which was initially expressed in stamen primordia and later in the tapetum during anther maturation. In agreement with its stage-specific expression pattern, the loss of function of the predominantly expressed B homeolog of AGO1d in a tetraploid durum wheat mutant resulted in smaller anthers with more infertile pollens than the wild type and a reduced grain number per spike. Together, our work provides a first glimpse of the gene regulatory networks in wheat inflorescence development that may be pivotal for floral and grain development, highlighting potential targets for genetic manipulation to improve future wheat yields.

Characterization of FLOWERING LOCUS T1 (FT1) gene in Brachypodium and wheat.

The phase transition from vegetative to reproductive growth is a critical event in the life cycle of flowering plants. FLOWERING LOCUS T (FT) plays a central role in the regulation of this transition by integrating signals from multiple flowering pathways in the leaves and transmitting them to the shoot apical meristem. In this study, we characterized FT homologs in the temperate grasses Brachypodium distachyon and polyploid wheat using transgenic and mutant approaches. Downregulation of FT1 by RNAi was associated with a significant downregulation of the FT-like genes FT2 and FT4 in Brachypodium and FT2 and FT5 in wheat. In a transgenic wheat line carrying a highly-expressed FT1 allele, FT2 and FT3 were upregulated under both long and short days. Overexpression of FT1 caused extremely early flowering during shoot regeneration in both Brachypodium and hexaploid wheat, and resulted in insufficient vegetative tissue to support the production of viable seeds. Downregulation of FT1 transcripts by RNA interference (RNAi) resulted in non-flowering Brachypodium plants and late flowering plants (2-4 weeks delay) in wheat. A similar delay in heading time was observed in tetraploid wheat plants carrying mutations for both FT-A1 and FT-B1. Plants homozygous only for mutations in FT-B1 flowered later than plants homozygous only for mutations in FT-A1, which corresponded with higher transcript levels of FT-B1 relative to FT-A1 in the early stages of development. Taken together, our data indicate that FT1 plays a critical role in the regulation of flowering in Brachypodium and wheat, and that this role is associated with the simultaneous regulation of other FT-like genes. The differential effects of mutations in FT-A1 and FT-B1 on wheat heading time suggest that different allelic combinations of FT1 homoeologs could be used to adjust wheat heading time to improve adaptation to changing environments.

[Influence of penetrative needling of Shendao (GV 11) on the symptom score and serum IgE content in chronic urticaria patients].

OBJECTIVE: To observe the changes of symptom scores and serum IgE level after treatment with thick-needle subcutaneous penetration of Shendao (GV 11) in chronic urticaria patients. METHODS: A total of 60 chronic urticaria patients were randomly divided into acupuncture group (n = 30) and medication group (n = 30). Subcutaneous penetrative needling was applied to GV 11 with thick acupuncture needle (retained for 4 h/time, once daily, 5 times/week) for patients of acupuncture group and Levocetirizine Hydrochloride tablets (5 mg/time, once daily in the first two weeks, then, once every other day in the 3rd and 41th weeks, and once every 3 days in the last two weeks) were given to patients of medication group. Serum IgE content was assayed before and 2,6, 12 weeks after the treatment by chemiluminescent technique. Symptom scores were obtained by "0-3 four levels assessment" method in the light of the size of the wheal and the itching severity. RESULTS: Self-comparison indicated that the symptom scores and serum IgE levels declined significantly (P < 0.01) 2 and 6 weeks (Wks) after the treatment in both acupuncture and medication groups,and 12 Wks after the treatment in the acupuncture group (P < 0.01). Comparison between two groups showed that the symptom score and serum IgE content of acupuncture group were significant lower than those of medication group 12 Wks after the treatment (P < 0.05). No significant differences were found between two groups in the symptom scores and serum IgE levels before, 2 and 6 Wks after the treatment (P > 0.05). A positive correlation exists between the symptom score and the serum IgE level before and after the treatment in both groups. CONCLUSION: Thick-needle subcutaneous penetration of Shendao (GV 11) can effectively improve clinical symptoms of chronic urticaria patients, which may be closely related to its effect in lowering peripheral blood IgE level.