Transcriptome Analysis Reveals the Mechanism of Exogenous Selenium in Alleviating Cadmium Stress in Purple Flowering Stalks (Brassica campestris var. purpuraria).

In China, cadmium (Cd) stress has a significant role in limiting the development and productivity of purple flowering stalks (Brassica campestris var. purpuraria). Exogenous selenium supplementation has been demonstrated in earlier research to mitigate the effects of Cd stress in a range of plant species; nevertheless, the physiological and molecular processes by which exogenous selenium increases vegetable shoots' resistance to Cd stress remain unclear. Purple flowering stalks (Brassica campestris var. purpuraria) were chosen as the study subject to examine the effects of treatment with sodium selenite (Na2SeO3) on the physiology and transcriptome alterations of cadmium stress. Purple flowering stalk leaves treated with exogenous selenium had higher glutathione content, photosynthetic capacity, and antioxidant enzyme activities compared to the leaves treated with Cd stress alone. Conversely, the contents of proline, soluble proteins, soluble sugars, malondialdehyde, and intercellular CO2 concentration tended to decrease. Transcriptome analysis revealed that 2643 differentially expressed genes (DEGs) were implicated in the response of exogenous selenium treatment to Cd stress. The metabolic pathways associated with flavonoid production, carotenoid synthesis, glutathione metabolism, and glucosinolate biosynthesis were among those enriched in these differentially expressed genes. Furthermore, we discovered DEGs connected to the production route of glucosinolates. This work sheds fresh light on how purple flowering stalks' tolerance to cadmium stress is improved by exogenous selenium.

Integrated network pharmacology to investigate the mechanism of Salvia miltiorrhiza Bunge in the treatment of myocardial infarction.

Salvia miltiorrhiza Bunge is a natural drug for treating myocardial infarction (MI). However, the targets and mechanisms of S. miltiorrhiza Bunge in the treatment of MI are yet to be elucidated. Traditional Chinese medicine systems pharmacology (TCMSP) data were used to screen out chemical constituents, and UniProt was used to predict relevant targets. Disease targets were obtained from the Online Mendelian Inheritance in Man and GeneCards databases. We used the STRING platform to build a protein-protein interaction network and used Cytoscape_v3.8.1 software to make a Drug-Ingredients-Gene Symbols-Disease network map. The Metascape database was used to perform gene ontology and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses for drug-disease overlapping gene symbols. The targets identified by network pharmacology were further verified by in vitro and in vivo experiments. Seventy-five active components of S. miltiorrhiza Bunge were obtained from the TCMSP database, while 370 disease targets and 29 cross-targets were obtained from the Genecards database. The KEGG pathway enrichment results suggested that the mechanism of S. miltiorrhiza Bunge in the treatment of MI was significantly related to the VEGF signalling pathway. In vitro and in vivo experiments were used to evaluate the reliability of some important active ingredients and targets. S. miltiorrhiza Bunge alleviated the damage to cardiac function, attenuated myocardial fibrosis and protected endothelial cell function by increasing the expression of TGF-ß and VEGFA. S. miltiorrhiza Bunge showed the therapeutic effect of MI by promoting the expression of VEGFA signalling pathway, providing a reliable basis for exploring herbal treatment of MI.

[Effects of acupuncture on carotid intima-media thickness and cerebral blood flow velocity in patients with mild-to-moderate hypertension].

OBJECTIVE: To observe the characteristics of carotid intima-media thickness (IMT) and cerebral blood flow velocity in patients with mild-to-moderate hypertension, and to evaluate the effects of acupuncture on carotid IMT and blood flow velocity of middle cerebral artery and vertebral-basilar artery. METHODS: A total of 240 patients with mild-to-moderate hypertension who met the inclusion criteria were treated with Huoxue Sanfeng acupuncture method proposed by academician SHI Xue-min. The acupoints of Renying (ST 9), Quchi (LI 11), Hegu (LI 4), Zusanli (ST 36) and Taichong (LR 3) were selected. The treatment was given once a day, five times a week for 3 months. The carotid ultrasonography and transcranial color Doppler were performed before treatment and 3 months after treatment to evaluate the improvements of carotid IMT and brain blood flow velocity. RESULTS: Among 175 patients, 94.3% suffered from impaired carotid IMT. After acupuncture intervention, 7.7%-10.9% patients had improved IMT but 4.6%-6.3% had aggravated carotid IMT. There was no significant difference of carotid IMT before and after treatment (P>0.05). About 50% patients had abnormal intracranial blood flow velocity; after acupuncture intervention, 27.4%-33.3% patients who had the abnormal blood flow velocity had normal one, but 27.0%-52.5% patients who had normal blood flow velocity had abnormal one. After acupuncture intervention, the low-speed blood flow of MCA, VA and BA in female patients aged 41-60 years and the low-speed blood flow of MCA and VA in female patients aged 61-70 years were significantly improved (all P<0.05); the high-speed blood flow of MCA and VA in male patients aged 61-70 years and the high-speed blood flow of VA and BA in female patients aged 41-60 years were significantly decreased (all P<0.05). CONCLUSION: Nearly 95% of patients with mild-to-moderate hypertension had carotid IMT, and about 50% had abnormal blood flow velocity of intracranial artery. The present study failed to found significant effects of acupuncture on carotid IMT, but it shows acupuncture can generally improve the low blood flow velocity in women with mild-to-moderate hypertension.

Effects of acupuncture with needle manipulation at different frequencies for patients with hypertension: Result of a 24- week clinical observation.

OBJECTIVES: To investigate whether the manipulation parameter of the twirling frequency in acupuncture affects the blood pressure when acupuncture is applied on the acupoints ST9 and LR3 in a human body. DESIGN: A randomized, controlled trial. INTERVENTIONS: A hundred and twenty patients with hypertension were randomized into four treatment groups. A twirling frequency of 120 twirls per minute or 60 twirls per minute were respectively applied on the acupoints ST9 or LR3. Each patient received five acupuncture sessions a week over a period of 12 weeks, with a follow-up period of a further12 weeks. The outcome was assessed by using an ambulatory blood pressure monitor. RESULTS: A hundred and twenty twirls per minute on ST9 could overall improve the BP (24hDBP, mSBP, mDBP, dDBP, nSBP and nDBP). Sixty twirls per minute on ST9 improved the 24hDBP, dSBP, dDBP and mDBP; 120 twirls per minute on LR3 did not show any anti-hypertensive effect, while 60 twirls per minute on LR3 improved the dSBP and dDBP. The results indicated that the acupuncture effect on BP, the onset of anti-hypertensive effect, the occurrence of the effect, and the duration of the effect depended on the frequency of needle manipulation on the same acupuncture point. CONCLUSIONS: As one of the important factors of the effects of acupuncture, the manipulation parameter's impact has its specificity for different acupoints. Thus, in the future, close attention needs to be paid to this clinically. TRIAL REGISTRATION: Chinese Clinical Trial Registry: Chi CTR-TRC-12002582.

The Role of Biologically Active Ingredients from Natural Drug Treatments for Arrhythmias in Different Mechanisms.

Arrhythmia is a disease that is caused by abnormal electrical activity in the heart rate or rhythm. It is the major cause of cardiovascular morbidity and mortality. Although several antiarrhythmic drugs have been used in clinic for decades, their application is often limited by their adverse effects. As a result, natural drugs, which have fewer side effects, are now being used to treat arrhythmias. We searched for all articles on the role of biologically active ingredients from natural drug treatments for arrhythmias in different mechanisms in PubMed. This study reviews 19 natural drug therapies, with 18 active ingredient therapies, such as alkaloids, flavonoids, saponins, quinones, and terpenes, and two kinds of traditional Chinese medicine compound (Wenxin-Keli and Shensongyangxin), all of which have been studied and reported as having antiarrhythmic effects. The primary focus is the proposed antiarrhythmic mechanism of each natural drug agent. Conclusion. We stress persistent vigilance on the part of the provider in discussing the use of natural drug agents to provide a solid theoretical foundation for further research on antiarrhythmia drugs.

A rapid and simple RP-HPLC method for quantification of kirenol in rat plasma after oral administration and its application to pharmacokinetic study.

A rapid and simple reverse-phase high-performance liquid chromatography (RP-HPLC) was developed and validated for the quantification of kirenol in rat plasma after oral administration. Kirenol and darutoside (internal standard, IS) were extracted from rat plasma using Cleanert™ C(18) solid-phase extraction (SPE) cartridge. Analysis of the extraction was performed on a Thermo ODS-2 Hypersil C(18) reversed-phase column with a gradient eluent composed of acetonitrile and 0.1% phosphoric acid. The flow rate was 1.0 mL/min and the detection wavelength was set at 215 nm. The calibration curve was linear over the range of 9.756-133.333 µg/mL (r(2) = 0.9991) in rat plasma. The lower limits of detection and quantification were 2.857 and 9.756 µg/mL, respectively. The intra- and inter-day precisions (relative standard deviation, RSD) were between 2.24 and 4.46%, with accuracies ranging from 91.80 to 102.74%. The extraction recovery ranged from 98.16 to 107.62% with RSD less than 4.81%. Stability studies showed that kirenol was stable in preparation and analytical process. The present method was successfully applied to the pharmacokinetic study of kirenol in male Sprague-Dawley rats after oral administration at a dose of 50 mg/kg.

Effects of selenium on the structure and function of recombinant human S-adenosyl-L-methionine dependent arsenic (+3 oxidation state) methyltransferase in E. coli.

The effects of Se(IV) on the structure and function of recombinant human arsenic (+3 oxidation state) methyltransferase (AS3MT) purified from the cytoplasm of Escherichia coli were studied. The coding region of human AS3MT complementary DNA was amplified from total RNA extracted from HepG2 cell by reverse transcription PCR. Soluble and active human AS3MT was expressed in the E. coli with a Trx fusion tag under a lower induction temperature of 25 degrees C. Spectra (UV-vis, circular dichroism, and fluorescence) were first used to probe the interaction of Se(IV) and recombinant human AS3MT and the structure-function relationship of the enzyme. The recombinant human AS3MT had a secondary structure of 29.0% alpha-helix, 23.9% beta-pleated sheet, 17.9% beta-turn, and 29.2% random coil. When Se(IV) was added, the content of the alpha-helix did not change, but that of the beta-pleated sheet increased remarkably in the conformation of recombinant human AS3MT. Se(IV) inhibited the enzymatic methylation of inorganic As(III) in a concentration-dependent manner. The IC(50) value for Se(IV) was 2.38 muM. Double-reciprocal (1/V vs. 1/[inorganic As(III)]) plots showed Se(IV) to be a noncompetitive inhibitor of the methylation of inorganic As(III) by recombinant human AS3MT with a K (i) value of 2.61 muM. We hypothesized that Se(IV) interacts with the sulfhydryl group of cysteine(s) in the structural residues rather than the cysteines of the active site (Cys156 and Cys206). When Se(IV) was combined with cysteine(s) in the structural residues, the conformation of recombinant human AS3MT changed and the enzymatic activity decreased. Considering the quenching of tryptophan fluorescence, Cys72 and/or Cys226 are deduced to be primary targets for Se(IV).

Structure-function roles of four cysteine residues in the human arsenic (+3 oxidation state) methyltransferase (hAS3MT) by site-directed mutagenesis.

Cysteine (Cys) residues are often crucial to the function and structure of proteins. Cys157 and Cys207 in recombinant mouse arsenic (+3 oxidation state) methyltransferase (AS3MT) are shown to be related to enzyme activity and considered to be the catalytic sites. The roles of some conserved Cys residues in the N-terminal region of the rat AS3MT also have been examined. However, little is known about the roles of the Cys residues in the middle region. The metabolism of inorganic arsenic in human is different from rat and mouse in some aspects though the AS3MT has a high degree of similarity in these species. In order to determine whether the Cys156 and Cys206 (corresponding to the catalytic sites, Cys157 and Cys207 in the mouse AS3MT) in the hAS3MT act as the catalytic sites and to study the roles of the Cys residues (Cys226 and Cys250) near the catalytic center in the middle region, we designed and prepared four mutants (C156S, C206S, C226S, and C250S) in which one Cys residue replaced by serine by PCR-based site-directed mutagenesis. The native form and cysteine/serine mutants were assayed for enzyme activity, free thiols, and the secondary structures by circular dichroism and Fourier transform infrared. Our data show that, besides C156S and C206S, C250S is another potential important site. C226S seems to have the same action as the wild-type hAS3MT with the consistent K(M) and V(max) values. Meanwhile, selenium can also inhibit the methylation of inorganic arsenic by C226S. All the mutants except C226S are calculated to have dramatic changes in the secondary structures. Cys250 might form an intramolecular disulfide bond with another Cys residue. These findings demonstrate that Cys residues at positions 156, 206, and 250 play important roles in the enzymatic function and structure of the hAS3MT.

[Progress of researches on pharmacokinetics of active components of Chinese drugs].

The pharmacokinetic research of Chinese drugs is still in the exploratory stage so far, a great progress has achieved in the researches on active components of them since the 1980s. The progresses in pharmacokinetic research of active components of some commonly used Chinese drugs were reviewed in this paper, and the problems to be solved in future were pointed out.

[A review of experimental designs of compound compatibility law of traditional Chinese medicine].

The paper discussed a variety of experimental designs of compound compatibility law of traditional Chinese medicine (TCM): study of whole formula and different ingredients of formula. The latter includes study of single ingredient, study of functional ingredient group, orthogonal design, clustering analysis, homogeneous design, factorial analysis and so on. It was proposed that experimental designs of formula should be based on the theory of TCM, and combined with modern sciences.