丝裂霉素C对膀胱肿瘤细胞株HSP70的诱导作用

目的研究丝裂霉素C对膀胱肿瘤细胞株TBC-1 HSP70的诱导作用。方法丝裂霉素C作用与TBC-1于2h后,观察在不同时间点0、8、24、48、72hTBC-1的HSP70的表达情况。结果丝裂霉素C化疗TBC-1后不同时间点0、8、24、48、72h的HSP70的表达OD值分别为0.306±0.005、0.255±0.009、0.203±0.018、0.237±0.029、0.239±0.020。正常TBC-1的HSP70表达OD值为0.247±0.022。结论丝裂霉素C可以诱导膀胱肿瘤细胞株TBC-1的HSP70的表达增高。     

卡介菌多糖核酸治疗充血性心力衰竭

目的 :研究卡介菌多糖核酸 (BCG PSN)对慢性充血性心力衰竭 (心衰 )病人免疫机能、心衰治疗周期及住院医疗费用的影响。方法 :5 9例心衰病人 ,常规抗感染、强心、利尿、扩血管等治疗 ,分BCG PSN组与常规组。BCG PSN组在常规治疗基础上给BCG PSN 1mg ,im ,qod× 2 6d。观察 2组T淋巴细胞CD4 /CD8,IgG ,IgA ,IgM及C3,住院费用及心衰缓解时间。另有 36例健康人作为对照。结果 :心衰病人IgG及C3升高 ,T细胞亚群CD4 /CD8下降 ;BCG PSN组较常规组治疗后IgG[(14 .2± 2 .4 ) g·L- 1vs (16 .3± 2 .4 ) g·L- 1,P <0 .0 5 ]和CD8[(30±5 ) %vs (38± 6 ) % ,P <0 .0 1]降低更明显 ,IgM[(3.0± 0 .6 ) g·L- 1vs (2 .1± 0 .5 ) g·L- 1,P <0 .0 1]及CD4 [(46± 6 ) %vs(38± 5 ) % ,P <0 .0 1]升高更明显 ;BCG PSN组较常规组心衰改善所需时间缩短 [(6± 3)dvs (8.4± 2 .6 )d ,P <0 .0 1],住院总药费有所降低。结论 :BCG PSN可提高心衰病人的免疫功能 ,加快心衰改善 ,住院费用降低     

重组hIFN-α-2b-卡介苗激活杀伤细胞抗膀胱肿瘤作用的体外研究

目的:研究重组hIFN-α-2b-卡介苗(hIFN-α-2b-BCG)对人外周血单个核细胞(peripheralbloodmonocytes,PBMC)活化增殖,以及BCG激活杀伤细胞(bacilliCalmette-Guérinactivatedkillercell,BAK细胞)对膀胱肿瘤细胞的杀伤效应。方法:分别利用野生型BCG、野生型BCG 干扰素作为对照诱导活化PBMC,比较重组BCG对PBMC的活化增殖效应,采用噻唑兰比色分析法(MTT法),分别在不同时间点检测上述各成分对PBMC增殖活化效果;采用乳酸脱氢酶释放试验(LDH法)研究不同浓度重组BCG活化PBMC后对膀胱肿瘤细胞的不同杀伤效果。结果:MTT测定表明,重组BCG诱导PBMC活化增殖的能力明显强于同浓度的对照组刺激效果(P<0.05),且重组BCG在72h仍保持较强的活化能力;乳酸脱氢酶释放试验显示,不同浓度重组BCG诱导的BAK细胞抗癌效应均高于对照组诱导的抗癌效果(P<0.05)。结论:重组BCG对人PBMC具有较强的活化增殖作用,增强PBMC对肿瘤细胞的杀伤效应。     

LW-AFC对环磷酰胺处理小鼠免疫功能的影响

目的探讨LW-AFC对免疫功能低下小鼠是否有改善作用。方法昆明小鼠ip给予环磷酰胺(CTX)0.02g·kg-1,连续10d,制备免疫功能低下小鼠模型。六味地黄(LW)浓缩丸(1.4g·kg-1)和LW-AFC(0.2,0.4和0.8g·kg-1)组小鼠在每次给予CTX后ig给予相应药物,连续10d。监测小鼠体重,以及胸腺和脾脏重量变化,并计算胸腺和脾脏系数;[3H]TdR掺入法检测脾淋巴细胞增殖反应,MTT法检测脾脏自然杀伤(NK)细胞杀伤活性,流式细胞术检测脾淋巴细胞中CD4+CD8-和CD4-CD8+T细胞亚群百分率,并计算CD4+/CD8+T细胞亚群比值。结果与正常对照组比较,模型组小鼠胸腺和脾重量及其系数明显降低,体重增长显著缓慢;LW和LW-AFC对小鼠体重增长缓慢、胸腺重量及其系数降低无明显改善作用;LW-AFC0.2g·kg-1对CTX导致的脾重及系数降低有明显改善(P<0.05)。模型组小鼠脾淋巴细胞自发增殖反应、刀豆蛋白A(ConA)和脂多糖(LPS)诱导的脾细胞增殖反应与正常对照组比较均明显降低,[3H]TdR掺入值由正常对照组的6115±441,19432±1778和(23345±7296)cpm分别降低为2741±340,9210±1387和(3983±263)cpm;与模型组比较,LW-AFC0.8g·kg-1对脾淋巴细胞自发增殖反应有明显的改善作用;LW浓缩丸和LW-AFC对ConA诱导的脾细胞增殖反应均具有明显的促进作用,[3H]TdR掺入值分别为13996±5161,27550±2356,15427±1444和(27333±1701)cpm;对LPS诱导的脾细胞增殖降低无改善作用,甚至低于CTX模型组(P<0.01)。模型组小鼠NK细胞杀伤活性由正常对照组的(39.5±0.5)%降低为(37.0±1.0)%,LW和LW-AFC明显促进小鼠NK细胞杀伤活性,杀伤率分别为(40.9±0.6)%,(39.7±0.8)%,(42.4±0.5)%和(39.8±0.9)%。与正常对照组比较,模型组小鼠脾脏CD3+,CD4+CD8-和CD4-CD8+T细胞百分率明显增加,CD4+/CD8+比值无明显变化;LW浓缩丸和LW-AFC可使CTX导致的升高的CD3+,CD4+CD8-和CD4-CD8+T细胞百分率明显降低,并可明显降低CD4+/CD8+比值。结论 LW-AFC对CTX所致小鼠免疫功能低下具有一定的改善作用。     

分枝杆菌和珊瑚红球菌细胞壁成分的免疫特性

前文报道了分枝杆菌(BCG)和珊瑚红球菌(Phodococcus corrallina,RC)细胞成分BCG-CWS和 RC-CWS的抗动物肿瘤作用。本文继续报道它们的免疫特性。 一 BCG-CWS和RC-CWS对PEC杀肿瘤细胞的影响。 BCG-CWS和RC-CWS油包水制剂,在50和100mcg/mouse剂量下,腹腔注射C57BL小鼠4天后,收集的PEC有明显的增强杀伤S180细胞的作用。其杀细胞指数分别为33.1±7.4,43.6±8.5,56.5±7.0和60.9±3.9。 二、BCG-CWS和 RC-CWS对淋巴细胞转化作用的影响。     

藻蓝蛋白亚基脂质体制备及其光动力学抗肿瘤作用实验

本研究建立了藻蓝蛋白亚基脂质体的制备方法，检测其对肿瘤细胞转染及光动力杀伤作用(PDT)效果。采用薄膜水合-超声分散法制备藻蓝蛋白亚基脂质体，测定其粒径大小及分布，荧光显微镜观察藻蓝蛋白亚基(PCS)及其脂质体(PCS-lip)的细胞转染率，MTT法检测藻蓝蛋白亚基及其脂质体对肿瘤细胞光动力杀伤效果。结果显示脂质体的粒径为80～160 nm，包封率为42.3%；在质量浓度为100 μg·mL^-1时，藻蓝蛋白亚基脂质体对小鼠肉瘤细胞S180转染率在2 h达到(18.5±0.8)%，4 h为(23.1±0.9)%，5～6 h转染率不再上升。肿瘤细胞光动力杀伤实验表明，在0～200 μg·mL^-1内，光照剂量为22 J·cm^-2时，随着藻蓝蛋白亚基及其脂质体浓度的增加，对胃癌细胞BGC-823和S180的光敏作用也随之增强；在200 μg·mL^-1时PCS-lip对两种细胞生存率分别降低为(45±5.2)%和(36±5.5)%，PCS-lip-PDT组与PCS-PDT组对细胞生存率影响有显著性差异(P<0.05)。研究结果表明，藻蓝蛋白亚基脂质体可很好地保持藻蓝蛋白亚基的生物活性，在相同蛋白浓度时藻蓝蛋白亚基脂质体比藻蓝蛋白亚基更快地转染进肿瘤细胞，药物作用的最佳时间为4 h。在相同浓度及光照剂量的情况下，藻蓝蛋白亚基脂质体对肿瘤细胞的光动力学作用略强于藻蓝蛋白亚基。     

熊去氧胆酸选择性诱导人肝肿瘤细胞凋亡及抑制增殖的实验研究

目的: 探讨熊去氧胆酸 (UDCA)对肝肿瘤细胞株诱导凋亡及抑制增殖的作用和机制。方法:用四氮唑蓝法、流式细胞术、TUNEL法、Wright Giemsa染色法、电镜及免疫细胞化学等方法,观察UDCA对肝肿瘤细胞株HepG2,BEL7402和正常人肝细胞株L 02的生长活力、细胞凋亡、细胞周期及Bax/bcl 2表达的影响。结果: UDCA对HepG2,BEL7402细胞株生长的抑制作用随药物浓度增高而增强(r2 =0. 96, P<0. 01;r2 =0. 97, P<0. 01;48h)。UDCA对HepG2及BEL7402的IC50分别为0. 92 mmol·L-1, 0. 86 mmol·L-1。UDCA(1. 0mmol·L-1 )对HepG2及BEL7402的凋亡率分别为(42±6) %及 (44±4) %,明显高于对照组(P<0. 01),并且阻滞细胞周期于S期。以UDCA(0. 8mmol·L-1 )处理HepG2 后bcl 2 表达由(24. 3±2. 4) %降低为 (10. 1±1. 6 ) %,Bax表达由(43±5) %升高为 (59±3) % (P<0. 01 );处理BEL7402细胞后bcl 2表达由 (21. 6±1. 8) %降为(11. 6±2. 1) %,Bax表达由 ( 44±4 ) %升高为(59±3) % (P<0. 01)。UDCA对L 02细胞无明显作用。结论: UDCA对HepG2, BEL7402细胞株有显著的抑制增殖及诱导凋亡作用,它可能与UDCA阻滞细胞周期、降低bcl 2和提升Bax的表达有关。     

超抗原SEC抗胶质瘤的研究

目的　观察超抗原SEC活化淋巴细胞对胶质瘤的杀伤作用。方法　采用APAAP法对SEC活化的淋巴细胞进行亚群分析 ;按MTT法测定经SEC活化的淋巴细胞对胶质瘤的体外杀伤作用 ;通过建立荷瘤动物模型及人外周血淋巴细胞———SCID小鼠免疫嵌合模型 ,观察肿瘤生长曲线。结果　经SEC活化的淋巴细胞主要为CD4 + 细胞 ;经SEC活化的淋巴细胞对 8株胶质瘤细胞均产生强大的杀伤作用 ,其中以 1× 10 4U·L-1剂量组作用最明显。生长曲线显示 :A组、C组和B组对胶质瘤生长抑制率分别为 5 8 5 %、4 6 2 %及 36 3%。结论　经超抗原SEC活化的淋巴细胞对胶质瘤有杀伤作用。     

树突状细胞以两种不同方式负载大肠癌细胞株体外刺激淋巴细胞抗肿瘤活性的比较

目的 比较树突状细胞(DC)以两种不同方式负载大肠癌细胞株体外刺激淋巴细胞的抗瘤活性.方法 分离正常人外周血单核细胞体外诱导DC,分别负载热休克诱导凋亡的大肠癌细胞株和肿瘤细胞裂解物,以此刺激淋巴细胞作为效应细胞,大肠癌细胞株为靶细胞.MTT法测定效应细胞对靶细胞的杀伤作用.结果 负载热休克大肠癌细胞的DC 与负载肿瘤细胞裂解物的DC都显示对靶细胞的杀伤活性,但前者的杀伤率明显高于后者(P＜0.05).结论 负载热休克肿瘤细胞的DC是一更为有效的负载方式.     

多沙唑嗪对映体对兔离体膀胱逼尿肌的作用及机制

目的观察多沙唑嗪(rac-DOX)及其对映体(S-DOX、R-DOX)对兔离体膀胱逼尿肌的作用并分析其机制。方法制备兔背侧和腹侧膀胱逼尿肌标本,记录药物或神经刺激诱发的膀胱平滑肌收缩反应。结果在兔背侧和腹侧膀胱逼尿肌标本,卡巴胆碱产生浓度依赖性收缩反应,两种标本的收缩反应差异无显著性(P>0·05)。在背侧膀胱逼尿肌标本,苯肾上腺素产生浓度依赖性收缩反应;但是,苯肾上腺素对腹侧膀胱逼尿肌无作用。S-DOX、R-DOX和rac-DOX在1μmol·L-1时,均可竞争性拮抗苯肾上腺素诱发的兔背侧膀胱逼尿肌收缩反应,其pKB值分别为7·44±0·19、7·39±0·14和7·38±0·30,三者的pKB值相同。电场刺激诱发兔背侧和腹侧膀胱逼尿肌产生稳定的收缩反应,该收缩反应被0·3μmol·L-1浓度的河豚毒素完全阻断。S-DOX、R-DOX和rac-DOX均抑制电场刺激诱发的背侧膀胱逼尿肌收缩反应(P<0·01),且三者的抑制作用强度差异无显著性(P>0·05);但是,它们对电场刺激诱发的腹侧膀胱逼尿肌收缩反应无影响。结论在兔离体膀胱背侧逼尿肌,S-DOX拮抗苯肾上腺素诱发收缩反应的pKB值与rac-DOX和R-DOX相同,三者尚能抑制电场刺激诱发的神经源性收缩反应。     

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.     

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.     

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.