SAR-guided development of indole-matrine hybrids as potential anticancer agents via mitochondrial stress/cytochrome c/caspase 3 signaling pathway.

Matrine is a clinically used adjuvant anticancer drug, yet its mild potency limited its application. To improve the anticancer activity of matrine, a total of 31 indole-matrine hybrids were constructed in four rounds of SAR-guided iterative structural optimization process. All of the synthesized compounds were evaluated for their antiproliferative activities against a panel of four human cancer cell lines (Hela, MCF-7, SGC-7901, HepG2) and two normal cell lines (GES-1, LO2). The most active hybrid 8g exhibited the anticancer IC50 values of 0.9 to 1.2 µM, which was 3-magnitude of orders more potent than matrine. 8g also showed better selectivity towards cancer cells with the selectivity index value raised from 1.5 to 6.2. Mechanistic studies demonstrated a mitochondrial distribution for 8g by intracellular click chemistry approaches, which led to the discovery that 8g strongly induced mitochondrial stress, as evidenced by impaired energy metabolism, depolarized mitochondrial membrane potential, overload of mitochondrial calcium and escalated ROS production. 8g-induced mitochondrial stress further led to the release of cytochrome c and subsequent activation of caspase 3, which significantly promoted cellular death and inhibited colony formation.

Antidepressant-like effect and phytochemical profile of supercritical CO2 extract from Citri reticulatae pericarpium.

Citri reticulatae pericarpium is a condiment, adding much flavor in Chinese food. Also it can be used to treat depression as a Traditional Chinese Medicine (TCM). The study here aimed to evaluate the antidepressant effect between the supercritical CO2 extract (SC-E) from Citri reticulatae pericarpium and the essential oil extracted by steam distillation (SD-E). And chemical compositions of SC-E were qualitatively analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) and gas chromatography-mass spectrometry (GC-MS). Compared with SD-E, SC-E showed a stronger antidepressant-like effect in FST and TST mice. And it also decreased the content of monoamine oxidase (MAO) in the cerebral cortex of stressed mice. A total of 60 compounds were identified in SC-E. Among them, 28 compounds were characterized in UPLC-Q-TOF/MS analysis and all are polymethoxyflavones (PMFs). Three main compounds, 3,5,6,7,8,3',4'-heptamethoxyflavone, nobiletin and tangeretin, together account for 66.09% of the total relative peak area. 33 terpenes were identified by GC-MS analysis, such as D-limonene (12.34%), ß-elemene (8.86%), germacrene D (5.59%) and (Z, E)-α-farnesene (5.44%). Polymethoflavones and terpenes are the main constituents of SC-E responsible for its antidepressant-like effect. The study could stimulate further investigations into the antidepressant effects and mechanism of Citri reticulatae pericarpium.

Neuroprotective Effect of Cyperi rhizome against Corticosterone-Induced PC12 Cell Injury via Suppression of Ca2+ Overloading.

Cyperi Rhizoma (CR) is a well-known functional food and traditional herbal medicine in Asian countries for the treatment of menstrual or emotional disturbances in women. Recent studies have shown the pharmacological effects of CR on neuronal diseases, such as Parkinson's disease (PD) and depression. Thus, the neuroprotective effect of CR might play a vital role in exerting its effect. Here, corticosterone-induced PC12 cells were applied to screen the active fraction of CR and evaluate its neuroprotective effect. The results indicated that the fraction containing medium-polarity chemical constituents (CR-50E) displayed the best protection effect. CR-50E could increase the cell viability and reduce cell apoptosis through inhibiting oxidative stress and decreasing the lactate dehydrogenase LDH release induced by corticosterone. Further, the mechanism of action was explored by cell metabolomics. The result showed CR-50E mediated the sphingolipids metabolism of corticosterone-induced PC12 cells, which suggested inhibition of Ca2+ overloading may involve the protection of CR-50E against cell damage. The expression levels of three key proteins in calcium transport, including phospholipase A2 (PLA2), calcium/calmodulin independent protein kinase II (CaMK II), and caspase-3, confirmed the above result by Western blot. The findings suggest that CR-50E can suppress the disequilibrium of calcium homeostasis-mediated apoptosis by improving the abnormal sphingolipids metabolism as well as remedying the damage of the cell membrane.

Rapid and visual detection of berberine hydrochloride based on a water-soluble pyrene derivative.

In this study, a rapid method for the detection of berberine hydrochloride (BRH) was developed based on a water-soluble pyrenyl probe, 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS). This method features low cost, good selectivity, high sensitivity and a fast response. The sensing mechanism of this probe is attributed to the formation of a complex between HPTS and BRH induced by electrostatic interaction and π-π stacking. To the best of our knowledge, this is the first fluorescent sensor for BRH based on organic materials that has low cost and a visual response. The detection limit of this method was as low as 1.24 µM and the linear response range is 2-50 µM. This method also allowed rapid detection of BRH real samples.

Metabolic profiling of hypoxia/reoxygenation injury in H9c2 cells reveals the accumulation of phytosphingosine and the vital role of Dan-Shen in Xin-Ke-Shu.

BACKGROUND: Xin-Ke-Shu (XKS), a patent medicine consisting of five commonly used traditional Chinese herbs, is used for the treatment of coronary heart diseases. A previous study showed that XKS has protective effects for ameliorating myocardial ischemia/reperfusion (I/R) injury. PURPOSE: This study was aimed to deeply understand the mechanisms and compatible principle of XKS against hypoxia/reoxygenation (H/R) injury and the contribution of each single herb to the efficacy of XKS. METHODS: An H/R model in H9c2 cardiomyocytes was applied to mimic I/R injury observed in vivo. The cell viability, the levels of LDH, MDA, SOD, and apoptosis were determined to evaluate the cardioprotection of XKS and its subtracted formula (knocked out one herb) in H/R injury. Cell metabolomics, combined with western blot analysis, was performed to uncover the inert molecular mechanism of XKS against H/R injury. RESULTS: Significant protective effects of XKS against oxidative stress and apoptosis induced by H/R injury were found in the pharmacodynamic evaluation. Moreover, the metabolic profile deviation of the H/R group from the control group was mainly ascribed to thirteen metabolites involved in four aberrant pathways, in which sphingolipid metabolism was revealed as the most relevant pathway involved in H/R injury (impact > 0.1). Notably, the accumulation of phytosphingosine (VIP = 5.84) was considered the most likely characteristic in H/R injury, which is well known to promote the opening of the mitochondrial permeability transition pore (mPTP) and activate cell apoptosis. Furthermore, XKS ameliorated all the abnormalities of the metabolic network in response to H/R injury. In agreement with this, a western blot analysis showed that XKS markedly regulated the over-expression of CaMK II and cleaved caspase-3. However, the subtracted formula showed no significant difference in comparison with the XKS group on protecting H/R injury except for QDS (subtracted Dan-Shen from XKS). CONCLUSION: The roots of Salvia miltiorrhiza Bge. (Dan-Shen) play an important role in the regulation of Ca2+ overloading, oxidative stress and apoptosis in H/R injury. Our study enabled information from holistic cell metabolomics to be used for mechanism and compatibility rule elucidations of TCMs.

Pharmacokinetics of costunolide and dehydrocostuslactone after oral administration of Radix aucklandiae extract in normal and gastric ulcer rats.

Costunolide and dehydrocostuslactone are the main active ingredients of Radix Aucklandiae (RA). An accurate and sensitive LC-MS/MS method was established to simultaneously determine contents of costunolide and dehydrocostuslactone in plasma. There were significant differences in pharmacokinetic parameters (AUC0-t, Cmax,1, Cmax,2, Tmax,1, Vd, and CL) of costunolide and dehydrocostuslactone between RA group and costunolide group or dehydrocostuslactone group. The relative bioavailability of costunolide or dehydrocostuslactone of RA extract was improved. As compared to normal group, the Tmax,2 values of dehydrocostuslactone of RA in gastric ulcer group were prolonged, while the Cmax,1, Cmax,2, and AUC0-t values decreased.

Probiotics administered intravaginally as a complementary therapy combined with antibiotics for the treatment of bacterial vaginosis: a systematic review protocol.

INTRODUCTION: Bacterial vaginosis (BV) is a highly prevalent vaginal polymicrobial disorder commonly encountered in women of childbearing age. Therapy with only recommended antibiotics results in low cure rates and unacceptably high recurrence rates. The use of probiotics as a complementary approach for use with antibiotics for the treatment of BV remains unclear. This review aims to assess the efficacy of lactobacilli administered intravaginally in conjunction with antibiotics for the treatment of BV. METHODS AND ANALYSIS: The Cochrane Central Register of Controlled Trials in The Cochrane Library, Cochrane Library of Systematic Reviews, Medline/PubMed and Embase will be used to search for articles from database inception to November 2016. Randomised controlled clinical trials using lactobacilli administered intravaginally in conjunction with antibiotics to treat BV will be included. Primary outcome will be the BV cure rate. The recurrence rate will be examined as secondary outcome. Two reviewers will independently select trials and extract data from the original publications. The risk of bias will be assessed according to the Cochrane Risk of Bias tool. We will perform data synthesis using the Review Manager (RevMan) software V.5.2.3. To assess heterogeneity, we will compute the I2 statistic. ETHICS AND DISSEMINATION: This study will be a review of published data and it is not necessary to obtain ethical approval. Findings of this systematic review will be published in a peer-reviewed journal. TRIAL REGISTRATION NUMBER: International Prospective Register of Systematic Reviews 2014: CRD42014015079.

Chaihu-Shu-Gan-San regulates phospholipids and bile acid metabolism against hepatic injury induced by chronic unpredictable stress in rat.

Chaihu-Shu-Gan-San (CSGS) is a famous classic traditional Chinese medicines (TCM) formula for treatment of liver stagnancy recorded in a famous book of traditional Chinese medicine, Jing Yue Quan Shu published in 1624. It has been extensively accepted as an antidepressant in China and its mechanism of action is still not clear. Previously we have found that hepatic injury happens in chronic unpredicted mild stress (CUMS). Thus, the protection of CSGS against hepatic injury induced by CUMS treatment was explored by metabonomics study and gene expression of the rat liver tissue. The results indicated that CSGS improved 8 of the 18 perturbed potential biomarkers in liver tissues of rats treated with CUMS, and involved in regulating phospholipids and bile acid metabolism against hepatic injury induced by CUMS in rat. The expressions of two apoptosis associated genes (Bcl-2 and Bax) and four genes (Pnpla6, Pla2g15, Baat and Gad1) related to the perturbed metabolic pathways were further investigated by quantitative real-time polymerase chain reaction (qRT-PCR). Both metabonomics and studies of genetic influences on metabolites demonstrated that CSGS inhibited hepatocyte apoptosis, and regulated phospholipids and bile acid metabolism against hepatic injury induced by CUMS in rat. Exploring the protection of CSGS against hepatic injury related to depression further clarify the relationship between CUMS-induced depression and hepatic injury, and also provide a novel insight to understand the underlying antidepressive mechanism of CSGS.

Acupuncture for benign prostatic hyperplasia: A systematic review and meta-analysis.

PURPOSE: This systematic review and meta-analysis aims to assess the therapeutic and adverse effects of acupuncture for benign prostatic hyperplasia (BPH) in randomized controlled trials (RCTs). METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, the Chinese Biomedical Database, the China National Knowledge Infrastructure, the VIP Database and the Wanfang Database. Parallel-group RCTs of acupuncture for men with symptomatic BPH were included. Data from the included trials were extracted by two independent reviewers and were analyzed with The Cochrane Collaboration Review Manager software (RevMan 5.3.5) after risk of bias judgments. The primary outcome measure of this review was a change in urological symptoms. RESULTS: Eight RCTs, which involved 661 men with BPH, were included. Follow-up varied from 4 weeks to 18 months. Pooling of the data from three trials that compared acupuncture with sham-acupuncture revealed that in the short term (4-6 weeks), acupuncture can significantly improve IPSS (MD -1.90, 95% CI -3.58 to -0.21). A sensitivity analysis of the short-term endpoint showed the same result (MD -3.01, 95% CI -5.19 to -0.84) with a borderline minimal clinical important difference (MCID). Qmax of the short-term endpoint indicated statistically positive beneficial effects of acupuncture (MD -1.78, 95%CI -3.43, -0.14). A meta-analysis after medium-term follow-up (12-18 weeks) indicated no significant effect on IPSS when the data from two trials were combined (MD -2.04, 95% CI -4.19, 0.10). CONCLUSION: Statistically significant changes were observed in favor of acupuncture in moderate to severe BPH with respect to short-term follow-up endpoints. The clinical significance of these changes needs to be tested by further studies with rigorous designs and longer follow-up times. TRIAL REGISTRATION NUMBER: PROSPERO CRD42014013645.

Effect of Different Solvents on the Measurement of Phenolics and the Antioxidant Activity of Mulberry (Morus atropurpurea Roxb.) with Accelerated Solvent Extraction.

The effects of 9 different solvents on the measurement of the total phenolics and antioxidant activities of mulberry fruits were studied using accelerated solvent extraction (ASE). Sixteen to 22 types of phenolics (flavonols, flavan-3-ols, flavanol, hydroxycinnamic acids, hydroxybenzoic acids, and stilbenes) from different mulberry extracts were characterized and quantified using HPLC-MS/MS. The principal component analysis (PCA) was used to determine the suitable solvents to distinguish between different classes of phenolics. Additionally, the phenolic extraction abilities of ASE and ultrasound-assisted extraction (UAE) were compared. The highest extraction efficiency could be achieved by using 50% acidified methanol (50MA) as ASE solvents with 15.14 mg/gallic acid equivalents g dry weight of mulberry fruit. The PCA results revealed that the 50MA followed by 50% acidified acetone (50AA) was the most efficient solvent for the extraction of phenolics, particularly flavonols (627.12 and 510.31 µg/g dry weight, respectively), while water (W) was not beneficial to the extraction of all categories of phenolics. Besides, the results of 3 antioxidant capability assays (DPPH, ABTS free radical-scavenging assay, and ferric-reducing antioxidant power assay) showed that water-based organic solvents increased the antioxidant capabilities of the extracts compared with water or pure organic solvents. ASE was more suitable for the extraction of phenolics than UAE.

Comparison of Polyphenol Profile and Inhibitory Activities Against Oxidation and α-Glucosidase in Mulberry (Genus Morus) Cultivars from China.

UNLABELLED: Mulberry (genus Morus) is a significant source of polyphenols, which can promote positive effects on human health. China has various mulberry cultivars, however, many Chinese mulberry cultivars have been only minimally studied. To solve this lack of research, 8 mulberry cultivars (Da10, Tang10, Yueshen74, Yuefenshen, Longsang, Ningxia1hao, Taiwanguosang, and Baiyuwang) from 4 regions of China were assessed to determine their polyphenol profiles using HPLC-MS/MS and then tested for their antioxidant and anti-α-glucosidase activities in vitro. A total of 18 nonanthocyanins and 4 anthocyanins were quantified in mulberry cultivars; among these polyphenols, chlorogenic acid, quercetin 3-O-rutinoside, and cyanidin 3-O-glucoside were confirmed as the major phenolic acid, flavonol derivative, and anthocyanin, respectively. Two types of stilbene compounds, piceid, and piceatannol, were detected for the 1st time in all mulberry cultivars. Moreover, the methanolic extracts of different mulberry cultivars showed disparate antioxidant and α-glucosidase inhibitory activities, and this discrepancy was mainly attributed to varying the anthocyanin content. Based on our results, Taiwanguosang is proposed to be a good candidate suitable for further process due to its high level of anthocyanins. PRACTICAL APPLICATION: The polyphenols of mulberry cultivars are vital for human health and are relevant to the further development of mulberry-based products. China has a wide range of mulberry cultivar resources, and many of these cultivars have not yet been studied. Our research concentrated on the polyphenol profiles, antioxidant, and α-glucosidase inhibitory activities of various mulberry cultivars from different regions of China to provide basic information for mulberry cultivar selection and mulberry-based food production.

Nutritional assessment of transgenic lysine-rich maize compared with conventional quality protein maize.

BACKGROUND: The gene sb401 encoding a lysine-rich protein has been successfully integrated into the genome of maize (Zea mays), its expression showing as increased levels of lysine and total protein in maize seeds. As part of a nutritional assessment of transgenic maize, nutritional composition, especially unintended changes in key nutrients such as proximates, amino acids, minerals and vitamins as well as in antinutrient (phytate phosphorus), and protein nutritional quality were compared between transgenic maize (inbred line 642 and hybrid line Y642) and conventional quality protein maize (QPM) Nongda 108. RESULTS: The contents of total protein, lysine, some other amino acids, several minerals and vitamin B2 in transgenic inbred line 642 and hybrid line Y642 were significantly higher than those in conventional QPM. Water-soluble protein and G2-glutelin were significantly promoted in transgenic maize Y642. CONCLUSION: Insertion of the lysine-rich sb401 gene increased the total protein and lysine content of transgenic maize varieties, leading to an improved amino acid score and therefore an improvement in the nutritive value of maize.

Cassane-type diterpenes from the seeds of Caesalpinia minax with their antineoplastic activity.

Five new cassane-type diterpenes, neocaesalpin AA (1), neocaesalpin AB (2), neocaesalpin AC (3), neocaesalpin AD (4) and neocaesalpin AE (5), were isolated from Caesalpinia minax together with three known compounds, 12α-methoxyl,5α,14ß-dihydroxy-1α,6α,7ß-triacetoxycass-13(15)-en-16,12-olide (6), spirocaesalmin (7) and magnicaesalpin (8). Their structures were elucidated based on 1D and 2D NMR, MS and CD analyses. Compounds 1-6 were tested against Hela, HCT-8, HepG-2, MCF-7 and A549 cancer cells and showed moderate cytotoxicity with IC50 values from 18.4 to 83.9 µM.

Added inositol regulates invertase secretion and glucose-repressed SUC2 gene expression in Saccharomyces sp. W4.

The effect of inositol supplementation on glucose derepression, invertase secretion and SUC2 gene expression in Saccharomyces sp. W4 was studied. Invertase secretion was repressed, when the yeast cells, grown the synthetic medium without inositol (I(-) medium) contained more than 0.2% (w/v) initial concentration of glucose. However, in the same medium plus inositol (I(+) medium, inositol conc. 100 microg/100 ml), invertase secretion was repressed only at glucose concentrations higher than 2.0% (w/v). Results showed that secreted invertase activity increased only in the I+ medium, whereas intracellular invertase activity remained constant in both media during the cell, growth. The mRNA encoding secreted invertase was higher in the glucose-derepressed cells grown in the I(+) medium than in the glucose-repressed cells grown in the I(-) medium. Similarly, phosphatidylinositol (PI) content was significantly higher in the cells grown in the I(+) medium than in the I(-) medium. These results indicated that PI might be involved in the glucose derepression, invertase secretion and SUC2 gene expression at the transcriptional level in the yeast.