淫羊藿苷对体外在无地塞米松培养基中诱导培养大鼠骨髓基质细胞成骨性分化的影响

目的：探讨在无地塞米松（dexamethasone）时淫羊藿苷（icariin,ICA）对体外培养大鼠骨髓基质细胞成骨性分化的影响,并试图寻找一种新的或更强的促体外培养骨髓基质细胞成骨性分化的诱导剂。方法用1×10－5 mol·L －1淫羊藿苷或1×10－8 mol·L －1的地塞米松对大鼠骨髓基质细胞分别进行药物干预,细胞计数试剂盒（Cell Counting Kit-8,CCK-8）检测细胞增殖,碱性磷酸酶检测试剂盒检测碱性磷酸酶（alkaline phosphatase,ALP）活性,茜素红染色检测钙化结节数量,实时定量聚合酶链反应（RT-PCR）检测骨保护素（osteoprotegerin,OPG）、核因子-κB 受体活化因子配体（receptor activator of NF-κB ligand, RANKL）、骨形态发生蛋白（bone morphogenetic protein,BMP）的信使 RNA（mRNA）表达情况。结果1×10－5 mol·L －1淫羊藿苷在无地塞米松的培养基中能显著提高大鼠骨髓基质细胞 ALP 活性;明显增加大鼠骨髓基质细胞钙化结节数目;显著上调BMP、OPG mRNA 水平以及 OPG／RANKL mRNA 比值。结论淫羊藿苷可能是一种潜在的促体外培养大鼠骨髓基质细胞成骨性分化的新诱导剂。     

脱水淫羊藿素与山柰素对体外培养成骨细胞成熟矿化影响的比较研究

比较研究脱水淫羊藿素与山柰素对体外培养大鼠成骨细胞(rat osteoblasts,ROB)增殖与成熟矿化的影响。取新生SD大鼠颅骨,多次酶消化法获得成骨细胞,培养于含10%FBS的MEM培养液中,3天后首次换液,铺满皿底90%以上后传代培养。分别用1×10 4、1×10 5、1×10 6和1×10 7mol.L 1的脱水淫羊藿素和山柰素进行药物干预,增殖分析采用MTT法,诱导培养9天后检测碱性磷酸酶(alkaline phosphatase,ALP)活性,筛选出最佳浓度。采用最佳浓度的脱水淫羊藿素和山柰素进行药物干预,比较脱水淫羊藿素、山柰素和对照组之间的ALP、钙盐沉积量、骨钙素分泌量、CFU-FALP和矿化结节数量等成骨性指标。提取总RNA,实时荧光定量PCR(real time PCR)检测BMP-2、Osterix(OSX)及Runx-2的mRNA表达情况,同时提取总蛋白,Western blotting法检测Ⅰ型胶原蛋白的分泌量。结果表明,1×10 4mol.L 1脱水淫羊藿素和山柰素对细胞增殖有抑制作用;1×10 5mol.L 1脱水淫羊藿素虽对ROB增殖无明显影响,但能显著提高成骨细胞中ALP活性、钙盐沉积量和骨钙素分泌量,增加CFU-FALP和矿化结节数量,与成骨细胞成熟矿化相关的因子BMP-2、OSX和Runx-2的基因表达量也显著升高,同时I型胶原蛋白的分泌量也明显增多;而山柰素并没有表现出促进体外培养ROB成熟矿化的特性。脱水淫羊藿素促进成骨细胞成熟矿化的作用明显高于山柰素,很可能是由脱水淫羊藿素8位C原子上存在的异戊烯基引起的。     

淫羊藿苷对乙酰胆碱酯酶的抑制作用

乙酰胆碱酯酶(acetylcholinesterase,AChE)抑制剂是目前治疗阿尔茨海默病(Alzheimer’s disease,AD)的主要药物。本文应用酶抑制动力学方法研究了淫羊藿苷对AChE的抑制作用,并利用分子荧光光谱法和分子对接分别研究了淫羊藿苷与AChE之间的结合反应和结合位点。研究发现,淫羊藿苷对AChE的活性具有一定的抑制作用,测得的IC50为3.50×10 8mol.L 1(他克林的IC50测得值为0.75×10 8mol.L 1)。淫羊藿苷对AChE的抑制作用表现为混合型可逆抑制,抑制常数KI和KIS分别为2.67×10 8和4.43×10 8mol.L 1。淫羊藿苷主要通过氢键作用和分子间作用力结合于AChE的外周阴离子位点。     

淫羊藿苷与金雀异黄酮骨保护作用的比较研究

目的通过淫羊藿苷(icariin,ICA)和金雀异黄酮(genistein,GEN)对幼龄大鼠和切除卵巢大鼠的药物干预,比较其抗骨质疏松的药理活性。方法 1月龄♀SD大鼠灌服25 mg·kg-1·d-1淫羊藿苷和10 mg·kg-1·d-1金雀异黄酮3个月,6月龄SD切除卵巢大鼠灌服同样的剂量3个月,检测骨密度、股骨生物力学、血清骨钙素与抗酒石酸性磷酸酶5b和骨组织形态。结果与对照组相比较,1月龄大鼠口服淫羊藿苷后,骨密度、股骨生物力学和骨质量均增加,而金雀异黄酮效果甚微。然而,切除卵巢大鼠在口服金雀异黄酮后,比淫羊藿苷更有效的抑制了骨量丢失和骨组织微结构的退化。结论相比较金雀异黄酮,淫羊藿苷有更强的成骨活性,但雌激素活性较弱,能更强地提高幼鼠峰值骨量。切除卵巢大鼠内环境由雌激素主导,因此,金雀异黄酮比淫羊藿苷能更强地减缓骨量丢失。     

淫羊藿苷调控mTOR/Akt/CREB通路对高糖诱导的足细胞自噬及凋亡的影响

目的 探讨淫羊藿苷对高糖诱导的足细胞自噬、凋亡及哺乳动物雷帕霉素靶蛋白(mTOR)/丝氨酸苏氨酸蛋白激酶(Akt)/环磷酸腺苷反应元件结合蛋白(CREB)通路的影响。方法 将小鼠足细胞MPC5分为5组：正常对照组(5.5 mmol·L^-1葡萄糖)、高糖组(30 mmol·L^-1葡萄糖)、淫羊藿苷组(30 mmol·L^-1葡萄糖+5μmol·L^-1淫羊藿苷)、GDC-0349组(30 mmol·L^-1葡萄糖+50μmol·L^-1 GDC-0349)、淫羊藿苷+GDC-0349组(30 mmol·L^-1葡萄糖+5μmol·L^-1淫羊藿苷+50μmol·L^-1 GDC-0349)。培养48 h后，噻唑蓝法检测MPC5细胞活力；吖啶橙染色观察MPC5细胞自噬情况；流式细胞术检测MPC5细胞凋亡；蛋白印迹法检测MPC5细胞自噬[微管相关蛋白1轻链3(LC3)Ⅱ、LC3Ⅰ、自噬相关蛋白(Beclin-1...     

HPLC法测定复方鹿茸酒中尿囊素、山药素Ⅰ、淫羊藿苷、淫羊藿次苷Ⅰ和宝藿苷Ⅰ

目的建立HPLC梯度洗脱法测定复方鹿茸酒中尿囊素、山药素I、淫羊藿苷、淫羊藿次苷I和宝藿苷I。方法采用Agilent TC-C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相A:甲醇–乙腈(1∶1)、流动相B:水,梯度洗脱;0~22 min在224 nm波长下检测尿囊素,22~45 min时在270 nm波长下检测山药素Ⅰ、淫羊藿苷、淫羊藿次苷Ⅰ和宝藿苷Ⅰ;体积流量0.9 m L/min;柱温30℃;进样量10μL。结果尿囊素、山药素I、淫羊藿苷、淫羊藿次苷I和宝藿苷I分别在7.98~159.60μg/m L、4.75~95.00μg/m L、9.18~183.60μg/m L、4.54~90.80μg/m L、4.92~98.40μg/m L与峰面积线性关系良好;平均回收率分别为98.67%、96.86%、99.56%、98.15%、97.90%,RSD值分别为1.12%、0.85%、1.07%、1.49%、0.97%。结论该方法简便、准确,重复性好,为进一步完善复方鹿茸酒的质量标准提供参考。     

淫羊藿次苷Ⅱ促进大鼠骨髓间充质干细胞成骨性分化过程中iNOS活性和NO生成量的变化

研究淫羊藿次苷II(icariside II,ICS II)对大鼠体外培养骨髓间充质干细胞(rat bone marrow stromal cells,rBMSCs)成骨性分化过程中诱导性一氧化氮合酶(induced nitric oxide synthase,iNOS)表达及NO生成的影响。贴壁筛选法体外培养rBMSCs,待铺满80%皿底时,进行成骨性诱导培养,同时采用1×10-5 mol.L-1 ICS II进行药物干预,比较ICS II组、L-NAME组、ICS II+L-NAME组和不加药的对照组之间的iNOS的活性、NO生成量,对比各组之间的成骨性指标,包括碱性磷酸酶活性、碱性磷酸酶阳性克隆数(CFU-FALP)及钙化结节数量。提取总RNA,实时荧光定量RCR(real-time PCR)检测Osterix(OSX)、Runx-2及iNOS mRNA的表达情况;同时提取总蛋白,Western blotting法检测Ⅰ型胶原蛋白和iNOS的分泌量。ICS II可显著增强碱性磷酸酶(ALP)活性,增加钙化结节和CFU-FALP数量,与成骨性分化相关的因子OSX和Runx-2的基因表达量也显著升高,同时I...     

基于模型群体分析的淫羊藿抗骨质疏松活性成分筛选研究

筛选淫羊藿中对破骨细胞生长具有抑制作用的活性成分。搜集不同产地淫羊藿药材,MTT法考察不同产地淫羊藿抗前破骨细胞RAW264.7生长活性。结果显示不同产地淫羊藿的抗RAW264.7生长活性具有显著差别。采用模型群体分析,一个具有潜在活性的化合物宝霍苷I被筛选出。通过破骨细胞活性验证,发现宝霍苷I活性优于淫羊藿苷。模型群体分析方法适用于指导中药活性成分预测及发现,为中药活性成分筛选提供了新方法。     

不同采收季节的淫羊藿中朝藿定C和淫羊藿苷的含量比较

目的比较3个贵州主流产淫羊藿药材在不同采收季节中朝藿定C和淫羊藿苷的含量。方法用高效液相色谱（HPLC）法测定粗毛淫羊藿、巫山淫羊藿、黔岭淫羊藿中朝藿定c和淫羊藿苷的含量。采用EliteSinoChromODS—AP柱（250mm×4．6mm,5．0Ixm）,流动相为乙腈（A）一水（B）梯度洗脱（0—22rain,27％_29％A;22～23rain,29％_÷100％A;23～34rain,100％A;34～36rain,loO％一27％A;36～50min,27％A）。结果朝藿定c和淫羊藿苷进样量分别在13．22～396．6Ixg（r=0．9999）和3．026～151．3斗g范围内与峰面积均呈良好线性关系（r=0．9999）,平均回收率均在98．0％一105．O％范围内;粗毛淫羊藿中朝藿定c和淫羊藿苷的含量分别为0．516％～2．973％,0．096％～0．216％;黔岭淫羊藿中朝藿定C和淫羊藿苷的含量分别为0．071％～0．185％,0．081％～0．164％;巫山淫羊藿含朝藿定c和淫羊藿苷中朝藿定c和淫羊藿苷的含量分别为1．015％～4．219％,0．080％～0．190％。结论巫山淫羊藿中朝藿定c和淫羊藿苷的含量最高,其次为粗毛淫羊藿、黔岭淫羊藿,说明不同产地、不同品种的淫羊藿药材中朝藿定c和淫羊藿苷的含量差异较大．     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Glycofection: The Ubiquitous Roles of Sugar Bound on Glycoplexes

Glycofection (transfection by using sugar-substituted polylysine) was assessed in order to provide an alternative to viral vectors for the transfer of genes into vascular smooth muscle cells. A rabbit vascular smooth muscle cell line (Rb-1 cells) was selectively transfected by using glycoplexes (glycosylated polylysine/pSV2LUC complexes) in the presence of 10 mu M of the fusogenic peptide GALA. A sugar-specific transfection was obtained when the glycofection was conducted for 1 h with glycoplexes containing either alpha Gal, alpha -Glc, alpha -GalNAc, beta -GlcNAc, or beta -GalNAc residues. The gene expression was high after transfection, with glycoplexes bearing alpha Gal, alpha -GalNAc, or beta -GalNAc residues that were weakly internalized, and low with glycoplexes carrying Lact or Rha residues that were well taken up by cells. These results suggest that 1) glycofection can be a good approach for a selective transfer of genes intovascular smooth muscle cells, 2) an efficient uptake of the glycoplexes is not the unique limiting step for an efficient transfection, and 3) the sugar-dependent trafficking of the glycoplexes inside the cells may account for the transfection efficiency.     

直肠癌下切缘病理组织学分析

目的探讨直肠癌逆向浸润与下切缘的安全距离的关系。方法对36例直肠癌Miles手术和Dixon手术后标本的肿瘤下缘1.0cm、2.0cm、3.0cm的肠壁及对应的系膜病理组织学检查,观察直肠癌逆向浸润或转移的距离。结果36例直肠癌标本距癌肿下缘1.0 cm、2.0cm、3.0cm的肠壁及对应的系膜病理组织学检查均为阴性,结论直肠癌远恻逆向浸润或转移未见超过1.0cm,因此认为保肛手术时切除肿瘤远侧肠管(包括系膜)2.0cm是安全的。