The effect of medicinal plants used in Chinese folk medicine on RANTES secretion by virus-infected human epithelial cells

The accumulation of inflammatory cells in the infective sites has been reported to play a crucial role in the progression of chronic inflammation and multiple sclerosis after viral infection. In the present study, nine ethanol extracts of Forsythia suspensa Vahl. (Oleaceae), Lonicera japonica Thunb. (Caprifoliaceae), Isatis indigotica Fort. (Cruciferae), Strobilanthes cusia (Ness.) O. Kuntze (Acanthaceae), Astragalus membranaceus (Fisch.) Bge. (Leguminosae), Hedysarum polybotrys Hand.-Mazz. (Leguminosae), Andrographis paniculata (Burm. f.) Ness. (Acanthaceae), Glycyrrhiza uralensis Fischer. (Leguminosae) and Ligusticum wallichii Franch. (Umbelliferae), medicinal plants traditionally used in China for treating conditions likely to be associated with inflammation and viral infection, were screened for their effect on RANTES secretion by influenza A virus (H1N1)-infected human bronchial epithelial cells (A549). With exception of Lonicera japonica, Isatis indigotica, Astragalus membranaceus and Hedysarum polybotrys, all plants tested at concentration of 200 microg/ml possessed more than 50% suppressing effect on RANTES secretion by H1N1-infected A549 bronchial epithelial cells. Among the plants tested, Andrographis paniculata showed the most promising property to inhibit RANTES secretion with an IC(50) of 1.2 +/- 0.4 microg/ml while the next two were Glycyrrhiza uralensis and Forsythia suspensa (IC(50) ranging from 35 to 48 microg/ml).     

Andrographolide prevents oxygen radical production by human neutrophils: possible mechanism(s) involved in its anti-inflammatory effect

We have reported that andrographolide (ANDRO), an active component of Andrographis paniculata, inhibits inflammatory responses by rat neutrophils. To further elucidate the possible mechanism(s) underlying the ANDRO's effect, N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced adhesion and transmigration of isolated peripheral human neutrophils were studied. Pretreatment with ANDRO (0.1 - 10 microM) concentration-dependently prevented fMLP-induced neutrophil adhesion and transmigration. We further examined the up-expression of surface Mac-1 (CD11b/CD18), an essential integrin mediated in neutrophil adhesion and transmigration. ANDRO pretreatment significantly decreased fMLP-induced up-expression of both CD11b and CD18. Accumulation of reactive oxygen species (ROS) as well as quick intracellular calcium ([Ca(++)](i)) mobilization induced by fMLP displays two important signalling pathways in regulating the up-expression of Mac-1 by neutrophils. That ANDRO pretreatment diminished fMLP-induced production of H(2)O(2) and O(2)*(-), but failed to block that of [Ca(++)](i) mobilization suggested that the ROS but not [Ca(++)](i) signalling could be modulated by ANDRO. To clarify whether ROS production impeded by ANDRO could be an antagonism of fMLP binding, phorbol-12-myristate-13-acetate (PMA), a direct protein kinase C (PKC) activator, was introduced to activate ROS production. PMA triggered remarkable ROS production and adhesion, and were partially reversed by ANDRO. This indicated that a PKC-dependent mechanism might be interfered by ANDRO. We conclude that the prevention of ROS production through, at least in part, modulation of PKC-dependent pathway could confer ANDRO the ability to down-regulate Mac-1 up-expression that is essential for neutrophil adhesion and transmigration.     

Pharmacological profile of evodiamine in isolated rabbit corpus cavernosum

This study was designed to examine the pharmacological properties of evodiamine in isolated rabbit corpus cavernosum. In phenylephrine-precontracted cavernosal strips, evodiamine (0.01-10 microM) induced a concentration-dependent relaxation. Endothelium removal, N(G)-nitro-L-arginine methyl ester (L-NAME), or 1-H-[1,2,4]oxadiazolo [4,3-alpha] quinoxalin-1-one (ODQ) treatment did not affect this effect. In endothelium-denuded preparations, evodiamine-evoked response was significantly reduced in 60 mM KCl-precontracted strips and by charybdotoxin treatment, but not by glibenclamide. Higher-concentration evodiamine (> or =10 microM)-induced relaxation was also accompanied by an increase in cAMP and cGMP levels, but this effect was not affected by cis-N-(2-phenylcyclopentyl)-azacyclotridec-1-en-2-amine mono-hydrochloride (MDL-12,330A, an adenylyl cyclase inhibitor) or ODQ (a guanylyl cyclase inhibitor), respectively. Evodiamine significantly augmented both the corporal relaxation and the accumulation of cyclic nucleotides to sodium nitroprusside and forskolin, respectively. Evodiamine also enhanced electrical field stimulation-evoked relaxation, and this additive effect was significantly counteracted by zaprinast. In preparations obtained from aged rabbits, evodiamine still elicited complete relaxation; in contrast, acetylcholine- and sodium nitroprusside-evoked maximal response was significantly blunted. In summary, evodiamine possesses a potent corporal relaxing effect which is attributable to endothelium-independent properties probably linked to charybdotoxin-sensitive K(+) channel activation in the cavernosal vasculature and by nonselective interfering phosphodiesterase to prevent cyclic nucleotide degradation. Furthermore, the physiological effects of evodiamine on the aged animals may implicate a potential for the treatment of erectile dysfunction.     

Antioxidant activity of piperlactam S: prevention of copper-induced LDL peroxidation and amelioration of free radical-induced oxidative stress of endothelial cells

The protective effects of piperlactam S, an alkaloid isolated from Piper kadsura (Choisy) Ohwi, on lipid peroxidation and free radical-mediated cell injuries were investigated. Piperlactam S (1 to 20 microM) concentration-dependently prevented the copper-catalyzed oxidative modification of human low-density lipoproteins (LDL) measured through (i) the lag period, (ii) the slope of the propagation phase, (iii) the total amount of conjugated dienes formed, and (iv) the electrophoretic mobility of LDL. Fe2+-induced oxidative modification of cell membrane was also significantly attenuated by piperlactam S as measured by thiobarbituric acid-reactive substances (TBARS). Furthermore, piperlactam S effectively minimized the loss of cell viability induced by Fenton's reagent (H2O2/FeSO4) in cultured endothelial cells and significantly reversed H2O2/FeSO4-induced impairment of endothelium-dependent relaxation to acetylcholine in rat aorta. Since the oxidative modification of LDL plays an important role in the genesis of atherosclerosis, piperlactam S may help to reduce the risk of atherosclerosis, not only by protecting LDL and membrane lipids from oxidative modification but also by reducing free radical-induced endothelial injury and/or dysfunction.     

Andrographolide suppresses the expression of inducible nitric oxide synthase in macrophage and restores the vasoconstriction in rat aorta treated with lipopolysaccharide

1. We investigated whether andrographolide, a diterpenoid lactone found at Andrographis paniculata, influences the induction of the inducible nitric oxide synthase (iNOS) in RAW264.7 cells activated by bacterial endotoxin (LPS), as well as in the rats with endotoxic shock and in aortic rings treated with LPS.
2. Incubation of RAW264.7 cells with andrographolide (1 to 50 μM) inhibited the LPS (1 μg ml⁻¹)-induced nitrite accumulation in concentration- and time-dependent manners. Maximum inhibition was observed when andrographolide was added together with LPS and decreased progressively as the interval between andrographolide and LPS was increased to 20 h.
3. Western blot analysis demonstrated that iNOS expression was markedly attenuated in the presence of andrographolide for 6–24 h, suggesting that andrographolide inhibited iNOS protein induction.
4. Thoracic aorta incubation with LPS (300 ng ml⁻¹) for 5 h in vitro exhibited a significant decrease in the maximal contractile response to phenylephrine (10⁻⁹–10⁻⁵ M). Andrographolide (30 μM) restored the contractile response to control level.
5. In anaesthetized rats, LPS (10 mg kg⁻¹, i.v.) caused a fall in mean arterial blood pressure (MAP) from 116±4 to 77±5 mmHg. The pressor effect of phenylephrine (10 μg ml⁻¹, i.v.) was also significantly reduced at 30, 60, 120 and 180 min after LPS injection. In contrast, animals pretreated with andrographolide (1 mg kg⁻¹, i.v., 20 min prior to LPS) maintained a significantly higher MAP when compared to LPS-rats given with vehicle. Administration of andrographolide 60 min after LPS caused a increase in MAP and significantly reversed the reduction of the pressor response to phenylephrine.
6. Our results indicated that andrographolide inhibits nitrite synthesis by suppressing expression of iNOS protein in vitro. And, this inhibition of iNOS synthesis may contribute to the beneficial haemodynamic effects of andrographolide in endotoxic shock.

     

ELSD-HPLC法监控维生素C生产过程

建立了ELSD-HPLC法监控由山梨醇生产维生素C的过程.采用氨基色谱柱,流动相为0.1%三氟乙酸-乙腈(梯度洗脱).4种成分均在0.025%～0.4%范围内线性关系良好.平均回收率分别为98.7%、100.5%、100.8%和100.4%.     

9-hydroxycanthin-6-one induces penile erection and delays ejaculation

IntroductionEurycoma longifolia Jack (Simaroubaceae) has the reputation as a male aphrodisiac because it is claimed to increase virility and sexual prowess. Nevertheless, whether or not E. longifolia regulates directly the muscle tone of corpus cavernosa and/or seminal vesicle (SV) remains unclear. Even until now, the compositions that could account for its aphrodisiac property are still unknownAimWe examined the effect of 9‐hydroxycanthin‐6‐one (9‐HC‐6‐one), a β‐carboline alkaloid isolated from E. longifolia, on penile erection and ejaculation, and further elucidated the mechanism of action.Main Outcome Measures9‐HC‐6‐one induces penile erection and delays ejaculation.MethodsDrug's effect was studied on rat corpus cavernosum (CC) and SV in vitro, and on the changes in intracavernosal pressure (ICP) after IC injection and intraluminal pressure (ILP) of the SV after hypogastric nerve stimulation (HNS), respectively.Results9‐HC‐6‐one relaxed significantly phenylephrine (PE)‐precontracted CC. Such response was not attenuated by endothelium disruption, N^G‐nitro‐L‐arginine methyl ester, or 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one treatment, suggesting that a nitric oxide/cyclic guanosine monophosphate‐dependent pathway was precluded. 9‐HC‐6‐one attenuated PE‐induced contraction by blocking cell surface and internal calcium channels with a higher potency for internal calcium release. This compound also antagonized calcium‐evoked contraction in Ca²⁺‐free, high K⁺‐depolarizing condition, suggesting that interfering with the entry of calcium through voltage‐dependent channels also contributed to 9‐HC‐6‐one‐induced corporal relaxation. After IC application of 9‐HC‐6‐one, a significant rise in ICP was observed as compared with the application of normal saline. 9‐HC‐6‐one relaxed significantly norepinephrine (NE)‐ and KCl‐precontracted SV, and antagonized NE‐induced oscillatory contraction as potent as clomipramine. Finally, the HNS‐evoked increase in ILP was dose‐dependently repressed after challenge by 9‐HC‐6‐one.Conclusion9‐HC‐6‐one might be the active component that contributed to the aphrodisiac effect of E. longifolia by antagonizing the smooth muscle tone of CC as well as SV probably through interfering with Ca²⁺ mobilization. Chiou W‐F and Wu T‐S. 9‐Hydroxycanthin‐6‐one induces penile erection and delays ejaculation. J Sex Med 12;9:1027–1036.     

高糖对人晶状体上皮细胞凋亡相关基因表达的影响

目的 通过检测高糖对人晶状体上皮细胞凋亡相关基因表达的影响,从基因角度探讨糖尿病患者白内障高发的机制.方法 体外培养人晶状体上皮细胞系,用不同浓度的葡萄糖孵育后,用四甲基偶氮唑盐比色法检测人晶状体上皮细胞活性,用荧光定量PCR方法检测多种凋亡相关基因的表达变化.结果 在以含不同浓度葡萄糖培养基培养细胞24 h后,发现晶状体上皮细胞活性在不同浓度的葡萄糖条件下产生明显变化,在葡萄糖浓度小于25.5 mmol·L-1时,晶状体上皮细胞增殖活力有少许提高,而当培养基中葡萄糖浓度大于35.5 mmol·L-1时,细胞增殖活力明显下降且呈刺量依赖性.同时该浓度下葡萄糖能够诱导多个凋亡相关基因高表达,其中凋亡基因p53和c-myc表达变化趋势与葡萄糖呈剂量依赖性,结果具有统计学意义(P＜0.05).结论 葡萄糖对人晶状体上皮细胞活性影响随浓度的改变而变化.高浓度下葡萄糖可通过诱导凋亡基因p53和C-myc高表达从而导致细胞凋亡.     

未成熟树突细胞诱导同种免疫低应答性的研究

目的：证实缺少辅助刺激分子的未成熟树突细胞（ＤＣ）在体外诱导同种Ｔ细胞的低应答性。方法：应用骨髓微血管内皮细胞作为饲养细胞扩增同品系小鼠骨髓细胞,经不同浓度ＧＭ－ＣＳＦ作用,在体外培养出不同发育阶段的ＤＣ。采用初次和再次单向混合淋巴细胞培养方法观察Ｂａｌｂ／ｃ小鼠脾脏Ｔ细胞对同种未成熟ＤＣ或脾细胞的应答能力。结果：在初次ＭＬＲ试验中,Ｂａｌｂ／ｃ Ｔ细胞对Ｃ５７ＢＬ／６小鼠未成熟ＤＣ刺激只呈现出微     

Concurrent chemoradiotherapy with/without induction chemotherapy in locoregionally advanced nasopharyngeal carcinoma: Long-term results of phase 3 randomized controlled trial

To report long-term results of a randomized controlled trial that compared cisplatin/fluorouracil/docetaxel (TPF) induction chemotherapy (IC) plus concurrent chemoradiotherapy (CCRT) with CCRT alone in locoregionally advanced nasopharyngeal carcinoma (NPC). Patients with stage III–IVB (except T3–4 N0) NPC were randomly assigned to receive IC plus CCRT (n = 241) or CCRT alone (n = 239). IC included three cycles of docetaxel (60 mg/m² d1), cisplatin (60 mg/m² d1), and fluorouracil (600 mg/m²/d civ d1–5) every 3 weeks. Patients from both groups received intensity-modulated radiotherapy concurrently with three cycles of 100 mg/m² cisplatin every 3 weeks. After a median follow-up of 71.5 months, the IC plus CCRT group showed significantly better 5-year failure-free survival (FFS, 77.4% vs. 66.4%, p = 0.019), overall survival (OS, 85.6% vs. 77.7%, p = 0.042), distant failure-free survival (88% vs. 79.8%, p = 0.030), and locoregional failure-free survival (90.7% vs. 83.8%, p = 0.044) compared to the CCRT alone group. Post hoc subgroup analyses revealed that beneficial effects on FFS were primarily observed in patients with N1, stage IVA, pretreatment lactate dehydrogenase ≥170 U/l, or pretreatment plasma Epstein–Barr virus DNA ≥6000 copies/mL. Two nomograms were further developed to predict the potential FFS and OS benefit of TPF IC. The incidence of grade 3 or 4 late toxicities was 8.8% (21/239) in the IC plus CCRT group and 9.2% (22/238) in the CCRT alone group. Long-term follow-up confirmed that TPF IC plus CCRT significantly improved survival in locoregionally advanced NPC with no marked increase in late toxicities and could be an option of treatment for these patients.