加味四逆散对皮质酮和谷氨酸致损伤的PCI2细胞中cAMP反应元件结合蛋白及其磷酸化的影响

目的研究中药复方加味四逆散（JWSNS）对皮质酮和谷氨酸致损伤的PCI2细胞中cAMP反应元件结合蛋白（CREB）及其磷酸化的影响。方法以200μmol·L^-1Cort和50μmol·L^-1Glu损伤的PC12细胞作为体外药理学研究模型,制备JWSNS含药血清,采用免疫组织化学法和Western blot法检测PC12细胞损伤模型中CREB和磷酸化CREB蛋白的表达。结果200μmol·L^-1 Cort和50Ixmol·L^-1 Glu可导致PCI2细胞中CREB和p-CREB表达下降（CREB阳性细胞率分别为17．1％±4．2％和20．8％±5．7％,p—CREB表达量分别为0．587±0．123和0．515±0．157,P＜0．05或P＜0．01）,10％JWSNS含药血清能升高CREB和P—CREB的表达（CREB阳性细胞率分别为62．6％±11．7％和79．5％±7．6％,p-CREB表达量分别为1．298±0．112和1．269±0．128,P＜0．05或P＜0．01）。结论10％JWSNS含药血清能通过调节cAMP—CREB信号通路发挥其神经保护和抗抑郁的作用。     

滇丹参对血管狭窄和金属基质蛋白酶的影响

目的 探讨滇丹参对高血脂兔腹主动脉球囊损伤后血管狭窄的影响，及其与基质金属蛋白酶、血管内膜中层平滑肌细胞增生的关系。方法 25只日本大耳兔随机分为正常对照组、球囊损伤组与滇丹参组。正常对照组不予任何方式处理。另两组行腹主动脉球囊损伤术，其中滇丹参组给予静脉注射滇丹参注射液（238mg·kg^-1·d^-1）2周，观察各组血管组织中基质金属蛋白酶表达情况的变化以及血管损伤处血管狭窄、血管内膜中层平滑肌细胞增生的情况。结果 应用滇丹参2周后滇丹参组血管组织中基质金属蛋白酶表达减少，血管损伤处血管内膜、外膜增生减轻，新生内膜面积减少，管腔面积增加。结论 滇丹参可以抑制血管组织中基质金属蛋白酶的表达，抑制血管内膜中膜平滑肌细胞增殖，抑制球囊损伤后兔腹主动脉血管的狭窄。     

丝裂素活化的蛋白激酶反义寡脱氧核苷酸防治血管内膜增殖

目的：探讨Ca^2 -钙调蛋白依赖性蛋白激酶（丝裂素活化的蛋白激酶）（CCDPK）在生长因子诱导体外培养大鼠血管平滑肌细胞增殖中的作用及反义CCDPK寡脱氧核苷酸（ODN）对球囊损伤后大白鼠血管内膜增生的抑制作用。方法：利用脂质体转染17－mer CCDPK反义ODN进入培养的血管平滑肌细胞以抑制CCDPK活性,设正义及随机ODN作对照。用蛋白质印迹法测定CCDPK表达。[^3H]胸腺嘧啶核苷酸掺入测定平滑肌细胞DNA合成。用2F球囊导管造成大白鼠颈动脉再狭窄模型,利用多聚胶F127－ODN系统由血管外膜部位给药。于损伤后2周取样,固定及HE染色观察内膜增生情况。FITC标记的ODN观察体内外给药方法的分布及吸收情况。结果：CCDPK反义ODN能明显抑制PDGF及ET诱导的CCDPK蛋白表达及[^3H]胸腺嘧啶核苷酸掺入。在大鼠颈动脉再狭窄模型,能明显抑制血管内膜增生。结论：CCDPK介导了PDGF及ET诱导的血管平滑肌细胞增殖。针对p42-和p44-CCDPK起始部位设计的17－mer反义ODN能有效抑制生长因子诱导的血管平滑肌细胞的增殖及球囊损伤大鼠血管内膜增生。     

大鼠胃腺癌发生中血管新生的研究

为研究大鼠胃腺癌发生中血管新生的情况,以 N-甲基-N~t-亚硝基-N~t-亚硝基胍(MNNG)诱发大鼠胃腺癌,于实验16、32周处死半数动物,处死前行活体心血管内墨汁灌注,进行增生细胞核抗原(PCNA)和血管内皮细胞生长因子(VEGF)免疫组化染色。结果显示,16周腺胃发生不典型增生病变,32周发展为原位癌和浸润癌。中、重度不典型增生病变处微血管密度为60.5±9.8,明显高于正常对照动物(42.3±8.1,P<0.01)。部分中、重度不典型增生及癌变区呈 VEGF 阳性反应。癌变区及中、重度不典型增生病变的 PCNA和微血管密度之间呈显著正相关(r=0.904,P<0.01)。上述结果表明,在胃癌发生过程中血管新生首先出现于不典型增生病变,与癌变关系密切;VEGF 可能参与癌前期病变的血管新生。     

Fas配体基因对大鼠颈动脉损伤后血管中层平滑肌细胞密度的影响

目的:探讨腺病毒载体介导Fas配体(FasL)基因导入对大鼠颈动脉损伤后血管中层平滑肌细胞密度的影响。方法:实验分治疗组(Ad-FasL组)和正常对照组两组(Ad-βgal组),每组动物均为6只。利用重组腺病毒载体分别将Ad-FasL、Ad-βgal导入大鼠颈动脉球囊损伤的内膜,3、14d后观察其对血管中层平滑肌细胞密度的影响。结果:与正常对照组相比较,通过腺病毒载体介导导入FasL基因的被球囊损伤的大鼠颈动脉,3d后损伤血管中层平滑肌细胞密度降低,14d后血管中层平滑肌细胞密度恢复到正常水平。结论:FasL可抑制血管中层平滑肌细胞密度,从而抑制球囊损伤后的内膜增生。     

大鼠实验性血栓模型的建立及其应用

目的：建立不同的动物血栓模型，评价阿司匹林的抗血栓作用。方法：根据文献建立角叉菜胶诱发小鼠尾动脉血栓模型、动脉埋线诱发大鼠颈动脉血栓模型以及大鼠血小板聚集的体外实验模型等，并进行改进。结果：角叉菜胶诱发小鼠尾动脉血栓形成，血栓发生率为90％，血栓长度为（15．8±1．9）mm，阿司匹林25～100mg·kg^-1，可以剂量依赖性降低血栓发生率和缩短血栓长度缩短。阿司匹林还可以剂量依赖性抑制动脉埋线诱发大鼠颈动脉血栓形成，血栓质量分别由（3．7±0．6）mg降低为（3．0±0．6），（2．3±1．3）。（1．8±0．4）mg，并能降低血浆中6-keto-PGF1／TXB2的比值。同时阿司匹林还可以剂量依赖性抑制大鼠体外血小板聚集率。结论阿司匹林具有确切的抗血栓形成作用。     

蝙蝠葛碱对大鼠血管内皮损伤后平滑肌细胞增殖的抑制作用

目的研究蝙蝠葛碱(Dau)对球囊损伤后血管平滑肌细胞(VSMC)增殖的抑制作用及其对细胞周期调控因子p27蛋白表达的影响。方法将25只♂SD大鼠随机均分为假手术组、单纯损伤组和Dau低、中、高剂量组,后4组行球囊内皮剥脱术,Dau组于术前3d开始,igDau25、50、100mg·kg-1·d-1,假手术组和单纯损伤组同期ig等量生理盐水。术后第14天处死所有大鼠,取胸主动脉进行HE染色,光镜下观察内膜损伤后形态学的变化,并进行图像分析。采用免疫组织化学技术检测球囊内皮损伤后血管平滑肌细胞p27蛋白表达的变化。结果球囊损伤后血管壁增生,Dau可剂量依赖性地减少新生内膜面积,增加管腔面积,降低球囊损伤后内膜及中膜厚度(P<0.05),抑制平滑肌细胞的增殖,促进p27蛋白的表达。结论p27参与了球囊损伤内皮后血管平滑肌细胞增殖的调节。Dau对损伤内皮所致平滑肌的增殖具有抑制作用,可防止血管再狭窄,其机理可能是通过调节p27的表达实现的。     

藻蓝蛋白抑制血管平滑肌细胞过度增殖性管腔狭窄的机制研究

目的联合体内外实验,探讨钝顶螺旋藻藻蓝蛋白(C-PC)抑制血管平滑肌细胞过度增殖、血管损伤后内膜增生和管腔狭窄的作用和机制。方法培养大鼠胸主动脉血管平滑肌细胞,用不同浓度的藻蓝蛋白进行干预,用氚标记胸腺嘧啶核苷(3H-TdR)参入率检测血管平滑肌细胞DNA合成和增殖状态,用Western blot和图像分析方法检测细胞周期蛋白依赖激酶抑制因子P21、P27的表达。构建大鼠胸主动脉球囊损伤后再狭窄动物模型,用藻蓝蛋白进行灌胃干预,术后14d取胸主动脉应用HE染色、免疫组化和计算机图像分析仪进行形态学、肌动蛋白(SM-α-actin)、增殖细胞核抗原(PCNA)、P21、P27表达水平检测。结果胎牛血清诱导的血管平滑肌细胞增殖活跃,P21、P27低表达;藻蓝蛋白明显促进P21、P27蛋白表达,明显抑制胎牛血清诱导的血管平滑肌细胞增殖和DNA合成。大鼠胸主动脉球囊损伤后,平滑肌细胞向内膜迁移、过度增殖、形成明显的新生内膜,导致管腔狭窄,SM-α-actin免疫组化证实新生内膜富含平滑肌细胞,PCNA高表达的同时P21、P27处于低表达水平;藻蓝蛋白干预后明显促进P21、P27基因表达,下调PCNA表达,抑制新生内膜形成、病理性血管重构,从而明显抑制管腔狭窄。结论钝顶螺旋藻藻蓝蛋白具有抑制血管平滑肌细胞过度增殖、血管损伤后管腔狭窄的作用,其机制与促进细胞周期关键调节因子P21、P27基因表达有关。     

骨髓低增生性病态造血的临床特点

为进一步探讨骨髓增生异常综合征（MDS）的临床特点,本文从患者初发病时的临床表现、血象、骨髓象、骨髓病理学、细胞遗传学等方面回顾性分析140例MDS的临床特点,并进行比较分析。结果表明,在低增生性MDS患者中,平均血红蛋白为（47．15&#177;15．63）g／L,平均血小板为（25．57&#177;18．84）&#215;10^9／L,平均中性粒细胞绝对值为（0．81&#177;0．58）&#215;10^9／L,染色体异常率30．77％。结论：低增生性MDS是MDS的一种特殊类型,不同于增生活跃／极度活跃性MDS。     

青霉素V钾咀嚼片的药动学和生物等效性研究

目的：研究青霉素V钾咀嚼片在健康人体的药物动力学及生物等效性。方法：19名健康志愿者随机双交叉、单剂量口服受试制剂和参比制剂0．472g，用液相色谱-串联质谱法（LC—MS／MS）测定人血浆中青霉素V的浓度，利用DAS2．0药物动力学软件计算有关药物动力学参数、相对生物利用度，并评价2种制剂的生物等效性。结果：受试制剂和参比制剂的tmax分别为（0．54&#177;0．09）h和（0．53&#177;0．13）h；Cmax分别为（10．64&#177;3．84）μg&#183;ml^-1和（10．87&#177;4．43）μg&#183;ml^-1；t1／2分别为（0．74&#177;0．20）h和（0．75&#177;0．10）h。AUC0-6h分别为（11．92&#177;4．04）μg&#183;ml^-1&#183;h和（12．34&#177;4．17）μg&#183;ml^-1&#183;h；AUC0-∞分别为（12．00&#177;4．04）和（12．43&#177;4．17）μg&#183;ml^-1&#183;h。青霉素V钾咀嚼片的相对生物利用度为（97．9&#177;12．8）％。结论：两种制剂具有生物等效性。     

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.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

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.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

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.