[Characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion based on consolidating the root and cultivating the primary].

Based on the theory of consolidating the root and cultivating the primary, the paper collates and reviews the theoretical evidences and the characteristics of Xin'an medical masters in treatment of bi syndrome with acupuncture and moxibustion so as to provide the ideas for further research. Xin'an medical masters thoroughly acquainted with the theory of consolidating the root and cultivating the primary in treatment of bi syndrome with acupuncture and moxibustion, emphasizing the regulation of qi and blood, yin and yang, the nutrient qi and the defensive qi; and replenishing the middle jiao (spleen and stomach) and the lower jiao (kidney). The acupoint selection was distinctive, in which, the acupoints were selected and stimulated in terms of the etiology and the pathogenesis of diseases, as well as the properties of special points. The remarkable therapeutic effect on bi syndrome was ensured through specially selecting he-sea points, ashi points and "yin-yang opposite" points; effectively using the penetrating needling technique and moxibustion method and combining acupuncture with herbal medication.

Direct Determination of Six Cytokinin Nucleotide Monophosphates in Coconut Flesh by Reversed-Phase Liquid Chromatography-Tandem Mass Spectrometry.

Coconut contains many uncharacterized cytokinins that have important physiological effects in plants and humans. In this work, a method based on liquid chromatography-tandem mass spectrometry was developed for identification and quantification of six cytokinin nucleotide monophosphates in coconut flesh. Excellent separation was achieved using a low-coverage C18 bonded-phase column with an acidic mobile phase, which greatly improved the retention of target compounds. To enable high-throughput analysis, a single-step solid-phase extraction using mixed-mode anion-exchange cartridges was employed for sample preparation. This proved to be an effective method to minimize matrix effects and ensure high selectivity. The limits of detection varied from 0.06 to 0.3 ng/mL, and the limits of quantification ranged from 0.2 to 1.0 ng/mL. The linearity was statistically verified over 2 orders of magnitude, giving a coefficient of determination (R2) greater than 0.9981. The mean recoveries were from 81 to 108%; the intraday precision (n = 6) was less than 11%; and the interday precision (n = 11) was within 14%. The developed method was applied to the determination of cytokinin nucleotide monophosphates in coconut flesh samples, and four of them were successfully identified and quantified. The results showed that trans-zeatin riboside-5'-monophosphate was the dominant cytokinin, with a concentration of 2.7-34.2 ng/g, followed by N6-isopentenyladenosine-5'-monophosphate (≤12.9 ng/g), while the concentrations of cis-zeatin riboside-5'-monophosphate and dihydrozeatin riboside-5'-monophosphate were less than 2.2 and 4.9 ng/g, respectively.

A novel method for the simultaneous analysis of seven biothiols in rice (Oryza sativa L.) using hydrophilic interaction chromatography coupled with electrospray tandem mass spectrometry.

Analysis of biothiols is still problematic, due to their high polarity, oxidation sensitivity and time-consuming sample preparation. In this paper, a direct, rapid and sensitive method was developed for simultaneous quantification of unbound cysteine (Cys), glutathione (GSH) and phytochelatins (PCs) in rice leaf, stem and root samples by hydrophilic interaction chromatography coupled with electrospray tandem mass spectrometry (HILIC-MS/MS). Homogenized samples were extracted with water containing 50mM dithiothreitol, without derivatization and further clean-up, and the extracts were injected directly onto an Xbridge Amide-HILIC column (3.5µm, 150mm×2.1mm i.d.). The best chromatographic separation and MS sensitivity was achieved using a linear gradient elution with 10mM aqueous ammonium formate and acetonitrile as the mobile phase. In MS/MS mode the detection limit (S/N≥3) of seven biothiols was 3-105nM. Good linearities were observed (r>0.995) with linear dynamic range at least over three orders of magnitude. Recoveries for most analytes were within the range of 77-128%, with relative standard deviations less than 18.2%. The intra-day precision (n=7) was 6.1-11.7%, and the inter-day precision over 15 d (n=15) was 8.5-16.3% for all biothiols. The optimized HILIC-MS/MS method was applied to study the influence of different cadmium (Cd) concentrations (0, 1 and 50µM) on contents of Cys, GSH and PC2-6 in rice tissue. With increasing Cd concentrations in nutrient solutions, contents of PC2-4 in rice roots increased but contents of Cys and GSH decreased. Contents of PC2-4 in both rice leafs and stems increased markedly at high dose Cd (50µM) treatment compared with controls, compared with low Cd concentrations (1µM). However, both PC5 and PC6 were not detected throughout the stress experiment.

[Construction of Saccharomyces cerevisiae whole-cell biocatalyst system for conversion miltiradiene].

Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.

[Maternal-neonatal vitamin D status and related factors].

OBJECTIVE: To determine the relationship between maternal and neonatal vitamin D status and related factors. METHOD: Serum 25-(OH)D levels were measured by ELISA in 499 pregnant women at 30 - 37 weeks gestation and in cord blood of their infants born at term (37 - 42 wk gestation) in Southeastern China at 28.9°N latitude. One-way analysis of variance (ANOVA) was used to explore maternal and neonatal vitamin D levels by season. Pearson linear and linear regression of partial correlation was used to analyze the relationship between maternal and neonatal 25-(OH) D levels. The multiple factors related to maternal vitamin D status was assessed by binary logistic regression. RESULT: The levels of serum 25-(OH)D were (33.0 ± 13.4) nmol/L in mothers and (31.0 ± 12.5) nmol/L in their newborns. Serum 25-(OH)D < 50 nmol/L was shown in 88.8% of mothers and 91.2% of their neonates. Both maternal and neonatal 25-(OH)D levels varied with season (Ps = 0.000). Vitamin D level was the lowest in spring, with the 25-(OH)D concentration < 50 nmol/L in 98.6% of mothers and 99.3% of their neonates. The highest vitamin D level was presented in fall, but there were still 64.0% of mothers and 75.0% of neonates with 25-(OH)D < 50 nmol/L. Except for season, calcium-vitamin D supplement and intake of egg ≥ 600 g per week during pregnancy benefited to improve maternal vitamin D level [25-(OH)D ≥ 50 nmol/L] [OR = 2.3 (95%CI:1.0, 5.3), 3.4 (95%CI:1.2, 9.9) respectively]. There was a positive correlation between maternal and neonatal 25-(OH)D measures in the sample as a whole (r = 0.45, P = 0.000, N = 499), the correlation was of no statistical significance when maternal serum 25-(OH)D was ≤ 25 nmol/L. CONCLUSION: Hypovitaminosis D was common in late pregnant mothers and their newborns in southeastern China, especially in spring. Vitamin D supplement and intake of vitamin D-rich food were beneficial to improvement of maternal vitamin D level. There was a moderate and positive correlation between maternal and neonatal 25-(OH)D concentrations in this population. The correlation was lost when maternal serum 25-(OH)D ≤ 25 nmol/L.