A novel NO-production-inhibiting triterpene and cytotoxicity of known alkaloids from Euonymus laxiflorus

A new triterpene, laxifolone A (1), four known sesquiterpene alkaloids, ebenifoline E-II (2), carigorinine E (3), euojaponine C (4), and emarginatine E (5), and six triterpenoids, 3-hydroxyolean-12-en-22,29-gamma-lactone, 3,11-dioxo-beta-amyrene, 3beta,22alpha-dihydroxyolean-12-en-29-oic acid, 28,29-dihydroxyfriedelan-3-one, 29-hydroxy-3-oxo-D:A-friedooleanan-28-oic acid, and putranjivadione, were isolated from the stems and leaves of Euonymus laxiflorus. Structural elucidations of these compounds were established by spectral analysis. Compound 1 displayed significant nitric oxide (NO) inhibitory effect.     

盐酸多柔比星脂质体注射液中磷脂膜成分的HPLC-ELSD测定

建立了HPLC-ELSD法测定盐酸多柔比星脂质体注射液磷脂膜中胆固醇、磷脂酰胆碱和溶血磷脂酰胆碱的含量,采用氨基色谱柱,流动相为2%草酸-乙醇-甲醇-氯仿(16:84:97:3),流速1ml/min.三者的平均回收率分别为98.8%、98.7%、98.6%,RSD为1.3%、0.9%、1.4%.     

Phenylpropanoid glycosides from Orobanche caerulescens

Two new phenylpropanoid glycosides, caerulescenoside ( 1), and 3'-methyl crenatoside ( 2), as well as five known phenylpropanoid glycosides [acteoside ( 3), isoacteoside ( 4), campneoside II ( 5), crenatoside ( 6), and desrhamnosyl acteoside ( 7)] were isolated from the whole plant of Orobanche caerulescens. The antioxidative effects of compounds 1 - 7 on human low-density lipoprotein were evaluated. All these compounds suppress concentration-dependently conjugated diene formation with IC (50) values of 1.25 +/- 0.06, 2.97 +/- 0.31, 0.31 +/- 0.01, 1.01 +/- 0.05, 1.15 +/- 0.04, 1.69 +/- 0.15, and 0.64 +/- 0.03 microM, respectively. Comparison of their antioxidative activities with that of resveratrol (IC (50) : 6.75 +/- 1.05 microM), a natural phenolic antioxidant isolated from grape, demonstrated that the prolonged effect on lag-time and the damping effect on oxidative rate by compounds 1 - 7 were all more potent.     

FGF-21 Plays a Crucial Role in the Glucose Uptake of Activated Monocytes

Monocytes display a gradual change in metabolism during inflammation. When activated, the increase in glucose utilization is important for monocytes to participate in immune and inflammatory responses. Further studies on the mechanism underlying this biological phenomenon may provide a new understanding of the relationship between immune response and metabolism. The THP-1 cells were used as a monocyte model. The cells were activated with lipopolysaccharide (LPS). Glucose uptake was measured using flow cytometry. The expression of fibroblast growth factor 21 (FGF-21), glucose transporter 1 (GLUT-1), and other FGF-21 signaling pathway-related factor mRNAs was determined by real-time polymerase chain reaction. Further, the relationship between FGF-21 expression in monocytes and phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt) signaling pathway was determined by Western blotting. LPS elevated FGF-21 expression in monocytic THP-1 cells in vitro. Functional assays showed that the phenomenon in which LPS and FGF-21 stimulated glucose uptake in monocytic THP-1 cells could be inhibited by FGFR inhibitor. The mechanism of elevation of FGF-21 was found to involve the PI3K/Akt signaling pathway. This study indicated that FGF-21 could regulate the immune response indirectly by influencing the glucose uptake of activated monocytes cells.     

Individualized induction chemotherapy by pre-treatment plasma Epstein-Barr viral DNA in advanced nasopharyngeal carcinoma

Background

The role of pretreatment Epstein-Barr virus DNA (pre-DNA) for individualized induction chemotherapy (IC) in locoregionally advanced nasopharyngeal carcinoma (LA-NPC) still remains unknown. We aimed to address this clinical issue.

Methods

In total, data on 6218 patient with newly diagnosed LA-NPC receiving concurrent chemoradiotherapy (CCRT) with or without IC were retrospectively reviewed. Receiver operating characteristics (ROC) curve was adopted to calculate the cut-off value of pre-DNA based on disease-free survival (DFS). Propensity score matching (PSM) method was adopted to balance prognostic factors and match patients. Survival outcomes between IC?+?CCRT and CCRT groups were compared.

Results

Among the original cohort, no survival difference between IC?+?CCRT and CCRT groups was found. The cut-off value of pre-DNA was 4650 copies/ml (area under curve [AUC], 0.620; sensitivity, 0.6224; specificity, 0.5673). For patients with Pre-DNA?≤?4650 copies/ml, the IC?+?CCRT and CCRT groups also achieved comparable survival outcomes (P?>?0.05 for all rates). However, IC?+?CCRT was associated with significantly improved 3-year DFS (78.6% vs. 74.8%, P?=?0.03), overall survival (OS; 91.4% vs. 87.5%, P?=?0.002) and distant metastasis-free survival (DMFS; 86.0% vs. 82.2%, P?=?0.036) for patient with pre-DNA?>?4650 copies/ml. Multivariate analysis also confirm that IC?+?CCRT was an independent prognostic factor for DFS (HR, 0.817; 95% CI, 0.683–0.977; P?=?0.027), OS (HR, 0.675; 95% CI, 0.537–0.848; P?=?0.001) and DMFS (HR, 0.782; 95% CI, 0.626–0.976; P?=?0.03).

Conclusions

Pre-DNA may be a feasible and powerful consideration for individualized IC apart from other baseline clinical characteristics in LA-NPC.

     

Relaxation of corpus cavernosum and raised intracavernous pressure by berberine in rabbit

1. In the present study, we have investigated the effect of berberine in rabbit isolated corpus cavernosum and measured the intracavernous pressure (ICP) change after intracavernosal injection of berberine in rabbit.
2. Berberine alone suppressed the basal tone and induced a concentration (0.1–100 μM)-dependent relaxation in phenylephrine (PE)-precontracted corpus cavernosum.
3. Tetrodotoxin (0.1 and 1 μM) treatment had no significant effect on the berberine-induced relaxation. Phentolamine (1 and 10 μM), propranolol (1 and 3 μM) and atropine (1 and 3 μM) were also without effect. These results suggest that berberine might cause relaxation of the cavernosal strip by direct action on the corpus cavernosum, not by a neuronal effect. Furthermore, muscarinic- and β-adrenoceptors were not involved.
4. Berberine-induced relaxations were significantly reduced by endothelium removal and by exposure to L-N^G-nitro arginine methyl ester (0.1 and 0.3 mM), but not indomethacin (30 μM).
5. In endothelium-deprived corpus cavernosal tissues, berberine-induced relaxations were significantly reduced in high K⁺ medium (KCl=60 mM), by charybdotoxin (ChTX) and 4-aminopyridine (4-AP) but not by glibenclamide and apamin.
6. After intracavernous injection of berberine (1, 2, 3 and 5 mg kg⁻¹), the ICP rose from 12.7±3.6 to 13.2±5.4, 25.3±6.1, 46.5±8.2, and 63.4±10.2 mmHg, respectively. The duration of tumescence ranged from 11.5–43.7 min.
7. The results show that berberine possesses a relaxant effect on rabbit corpus cavernosal tissues which is attributable to both endothelium-dependent and-independent properties. While the former component is apparently due to the release of NO from sinusoidal endothelium, the endothelium-independent mechanism involved in berberine relaxation is probably linked to ChTX- and 4-AP-sensitive K⁺ channel activation in the cavernosal vasculature.

     

筛选糖尿病候选药物FGF-21受体激动剂的新型细胞模型的建立

建立以成纤维细胞生长因子-21(fibroblast growth factor-21,FGF-21)信号通路为靶点的药物筛选细胞模型,用于筛选FGF-21受体激动剂类的新型治疗糖尿病药物。FGF-21的生理功能主要是通过细胞表面的FGFR及辅助受体βklotho传递信号,从而激活细胞内相关调控的一系列信号通路以及基因转录来实现的。本实验将βklotho基因构建到逆转录病毒表达载体pBMN-IRES-EGFP。将该载体导入包装细胞,收集病毒上清液并感染3T3-L1细胞,筛选到稳定表达βklotho细胞系。该细胞模型在FGF-21作用下可以促进细胞葡萄糖转运蛋白-1表达及转运水平升高,从而提高细胞糖吸收能力。构建该细胞系可以方便对FGF-21及类似药物进行高通量筛选,为糖尿病药物的研究提供了一种新方法。     

1,3,5-trihydroxy-4-prenylxanthone represses lipopolysaccharide-induced iNOS expression via impeding posttranslational modification of IRAK-1

Both high level of nitric oxide (NO) and its generating enzyme, inducible NO synthase (iNOS), play important roles in pathophysiological conditions such as inflammatory processes. We previously found that 1,3,5-trihydroxy-4-prenylxanthone (TH-4-PX) isolated from Cudrania cochinchinensis repressed lipopolysaccharide (LPS)-induced NO production in RAW264.7 macrophages. Here we further examined the underlying mechanisms using RT-PCR and Western blot analyses. Consistent with NO inhibition, suppression of LPS-induced iNOS expression by TH-4-PX through abolishing IκB kinase (IKK) phosphorylation, IκB degradation and nuclear factor-κB (NF-κB) nuclear translocation was observed. After LPS stimulation, the increased nuclear level of c-Fos and c-Jun (major components of activator protein-1, AP-1) and the phosphorylated level of upstream signal molecules, such as c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase, (ERK) were all significantly suppressed by TH-4-PX, while p38 remained unaffected. A further experiment revealed that TH-4-PX inhibited the phosphorylation of transforming growth factor-β (TGF-β)-activated kinase 1 (TAK1), an upstream signaling molecule required for IKK and mitogen-activated protein kinases (MAPKs) activation. Stimulation with LPS also triggered the modification (phosphorylation and ubiquitination) and eventually the proteasomal degradation of membrane-associated interleukin (IL)-1 receptor-associated serine/threonine kinase 1 (IRAK-1), an essential signaling component to toll-like receptor (TLR)-mediated TAK-1 activation. Interestingly, the modified pattern of IRAK-1 in the presence LPS was significantly attenuated by TH-4-PX treatment. In conclusion, TH-4-PX inhibited LPS-induced NF-κB and AP-1 activations by interfering with the posttranslational modification (phosphorylation and/or ubiquitinylation) of IRAK-1 in the cell membrane to impede TAK1-mediated activation of IKK and MAPKs signal transduction.