辛伐他汀对自体移植静脉内膜增生的抑制作用

目的 研究辛伐他汀对自体移植静脉内膜增生的影响.方法 48只健康成年家兔随机均分为辛伐他汀治疗组(A组)和生理盐水对照组(C组).建立自体静脉旁路移植模型,术后3、7、28 d切取移植静脉,观察组织病理变化,测量新生内膜厚度及面积,检测静脉壁增殖细胞核抗原(PCNA)表达,RT-PCR检测静脉壁基质金属蛋白酶(MMP)2、MMP-9 mRNA表达,明胶酶谱法检测MMP-2、MMP-9活性.结果 与C组比较,A组术后内膜增生、移植静脉壁阳性细胞数以及MMP-2、MMP-9 mRNA表达和活性均明显减少(P<0.05).结论 辛伐他汀可抑制自体移植静脉内膜增生,其作用机制可能是通过抑制血管平滑肌细胞(VSMC)增殖并限制其向内膜迁移而实现的.     

MMP-2、MMP-9在自体移植静脉表达及活性变化

目的 探讨自体移植静脉基质金属蛋白酶(MMP)-2和MMP-9表达及活性的变化.方法 建立兔自体静脉旁路移植模型后3、7、28 d切取移植静脉,取对侧未移植静脉为自身对照.行HE和Verhoeff弹性纤维染色观察组织病理变化,计算机病理图像分析系统测量新生内膜厚度及面积,半定量逆转录-PCR检测静脉壁MMP-2和MMP-9 mRNA表达,明胶酶谱法检测MMP-2、MMP-9活性,应用免疫组化法检测MMP-2蛋白表达.结果 术后7、28 d移植静脉内膜明显增生.与未移植静脉相比,术后3 d,移植静脉MMP-2、MMP-9 mRNA表达已明显增高,7 d达峰值(P＜0.01),术后28 d渐下降至正常水平,但MMP-9 mRNA表达仍维持较高水平.术后三个时间点移植静脉壁MMP-2、MMP-9活性显著增加.结论 自体移植静脉MMP-2、MMP-9表达及活性显著增强,促进移植静脉内膜增生.     

自拟川黄三七汤防治自体移植静脉再狭窄的实验研究

目的研究自拟川黄三七汤对兔颈外静脉移植至股动脉后再狭窄的防治作用,并探讨可能的机制。方法 50只新西兰大白兔,随机分为实验组(川黄三七组)、对照组(生理盐水组)。实验组术前3d开始采用灌胃法给予川黄三七汤,对照组给与等量生理盐水,1次/d,直至术后取材。所有动物于术日建立自体静脉旁路移植模型:利用显微外科技术将颈外静脉端侧吻合于股动脉,于术后4周取血管桥标本,同时取0.5cm移植静脉作为正常静脉参照。光镜下观察、测定移植静脉内膜中膜厚度变化。扫描电镜观察近吻合口处和中央部静脉桥的管腔狭窄程度,管腔增生面积与管腔面积比(S1/S2),统计学分析各组间差异。结果移植4周后,各组静脉均有内膜及中膜较正常颈静脉明显增厚表现,实验组内膜及中膜增厚较对照组轻。两组静脉桥的管腔狭窄程度进行比较存在明显差异。结论川黄三七汤能抑制动脉化静脉内膜及中膜增厚,有减轻桥静脉狭窄的作用。     

The Expression and Clinical Significance of PPAR-γ, p27kip1 and Skp2 in Hepatocellular Carcinoma

目的:探讨过氧化物酶增殖物激活受体(PPAR)-γ、p27kip1和细胞S期激酶相关蛋白(Skp)2蛋白在肝癌发生发展过程中的作用.方法:肝细胞癌组织标本57例(肝癌组)和正常肝组织标本(对照组)20例.采用免疫组织化学方法检测2组PPAR-γ、p27kip1及Skp2蛋白的表达,分析它们之间及其与肝癌临床病理特征的关系.结果:肝癌组PPAR-γ及Skp2蛋白的阳性表达率高于对照组,p27kipl蛋白阳性表达率低于对照组(P＜0.05),PPAR-γ蛋白与Skp2、P27kip1蛋白在不同TNM分期中的表达差异均有统计学意义(P＜0.05).PPAR-γ蛋白与Skp2、p27kip1蛋白表达无相关性(P＞0.05);Skp2与p27kip1蛋白的表达呈负相关(r=-0.307,P=0.020).Skp2蛋白低表达组的1、3、5年生存率高于高表达组(P＜0.001).结论:PPAR-γ和Skp2蛋白表达与肝细胞癌的恶性生物学行为密切相关,Skp2可作为判断肝细胞癌预后的指标.     

MMP-2、MMP-9在自体移植静脉的表达及活性变化

目的探讨自体移植静脉基质金属蛋白酶（MMP）-2和MMP-9表达及活性的变化。方法建立兔自体静脉旁路移植模型后3、7、28d切取移植静脉,取对侧未移植静脉为自身对照。行HE和Verhoeff弹性纤维染色观察组织病理变化,计算机病理图像分析系统测量新生内膜厚度及面积,半定量逆转录-PCR检测静脉壁MMP-2和MMP-9mRNA表达,明胶酶谱法检测MMP-2、MMP-9活性,应用免疫组化法检测MMP-2蛋白表达。结果术后7、28d移植静脉内膜明显增生。与未移植静脉相比,术后3d,移植静脉MMP-2、MMP-9mRNA表达已明显增高,7d达峰值（P〈0.01）,术后28d渐下降至正常水平,但MMP-9mRNA表达仍维持较高水平。术后三个时间点移植静脉壁MMP-2、MMP-9活性显著增加。结论自体移植静脉MMP-2、MMP-9表达及活性显著增强,促进移植静脉内膜增生。     

PPAR-γ和p27~(kip1)、Skp2在肝细胞癌中的表达及临床意义

目的:探讨过氧化物酶增殖物激活受体(PPAR)-γ、p27kip1和细胞S期激酶相关蛋白(Skp)2蛋白在肝癌发生发展过程中的作用。方法:肝细胞癌组织标本57例(肝癌组)和正常肝组织标本(对照组)20例。采用免疫组织化学方法检测2组PPAR-γ、p27kip1及Skp2蛋白的表达,分析它们之间及其与肝癌临床病理特征的关系。结果:肝癌组PPAR-γ及Skp2蛋白的阳性表达率高于对照组,p27kip1蛋白阳性表达率低于对照组(P0.05),PPAR-γ蛋白与Skp2、P27kip1蛋白在不同TNM分期中的表达差异均有统计学意义(P0.05)。PPAR-γ蛋白与Skp2、p27kip1蛋白表达无相关性(P0.05);Skp2与p27kip1蛋白的表达呈负相关(r=-0.307,P=0.020)。Skp2蛋白低表达组的1、3、5年生存率高于高表达组(P0.001)。结论:PPAR-γ和Skp2蛋白表达与肝细胞癌的恶性生物学行为密切相关,Skp2可作为判断肝细胞癌预后的指标。     

非小细胞肺癌中p27kip1、p16蛋白的表达及临床意义

目的探讨非小细胞肺癌中p27kip1、p16基因蛋白的表达及临床意义.方法采用免疫组化Envision法检测57例非小细胞肺癌(NSCLC)标本p27kip1、p16蛋白的表达.结果 (1)p27kip1、P16在非小细胞肺癌中阳性表达率分别为43.86%、49.12%,在癌旁正常支气管上皮阳性率分别为92.0%、96.0%,其在肺癌和正常组织中表达的差异有显著意义(P<0.005);(2)p27kip1、p16阳性表达者生存期分别较p27kip1、p16阴性表达者延长(P<0.05),且与淋巴结转移呈负相关(P<0.025);(3)p27kip1与p16的表达显著正相关(P<0.05).结论 p27kip1、p16低表达与非小细胞肺癌的发生、进展密切相关,与生存期呈负相关,两者可作为综合判断患者预后的指标.     

雷帕霉素通过mTORC1/p70S6K通路调控大鼠系膜细胞增殖及细胞周期的机制研究

为了探究雷帕霉素对大鼠系膜细胞增殖、细胞周期的影响及其作用机制,将大鼠系膜细胞(HBZY-1)分为以下6组:对照组(control)、血小板衍生生长因子(platelet derived growth factor, PDGF) 20 ng·mL-1组、PDGF+雷帕霉素1、10、100和1 000 nmol·L-1组; MTT法和流式细胞术检测雷帕霉素(1、10、100、1 000 nmol·L-1)作用后对细胞增殖和周期的影响; Western blot法检测周期蛋白D1 (cyclin D1)、周期蛋白E (cyclin E)、周期素依赖性蛋白激酶2(cyclin-dependent kinase 2, CDK2)、周期素依赖性蛋白激酶4 (cyclin-dependent kinase 4, CDK4)、p70S6K/p-p70S6K和p27的蛋白表达; Real-time-PCR法检测p27 mRNA。结果显示,雷帕霉素能显著抑制PDGF刺激系膜细胞(glomerular mesangial cells, MCs)引起的增殖,且呈剂量及时间依赖性,但随着浓度增加(1～1 00...     

雷帕霉素对肺间质纤维化大鼠IL-17、IL-2、TGF-β和IL-6及调节性T细胞的影响

目的 研究雷帕霉素对大鼠肺纤维化的影响及其机制.方法 SD大鼠15只,随机分为博来霉素5只,生理盐水对照组5只,雷帕霉素干预组5只.采用气管内注入博来霉素（5 mg·kg-1）的方法复制大鼠肺纤维化模型.对照组采用气管内注入生理盐水（1.25 mL·kg-1）的方法制造阴性对照.气管灌注博来霉素后第3天给予雷帕霉素干预组大鼠雷帕霉素灌胃（每天1 mg·kg-1）,连续10 d达到雷帕霉素对肺纤维化的干预.第28天分别处死动物,肺组织病理切片HE染色,免疫组织化学方法检测IL-17、IL-2、TGF-β和IL-6的表达;流式细胞术检测CD4＋调节性T细胞和CD8＋调节性T细胞的比例.结果 雷帕霉素干预组肺纤维化严重程度较博来霉素组明显减轻,Ashcroft评分存在统计学差异（P＜0.01）; IL-6、IL-17、TGF-β和IL-2表达水平在博来霉素组较正常对照组和雷帕霉素干预组升高（P＜0.01）.雷帕霉素组的IL-17、TGF-β的表达水平较对照组升高（P＜0.01）,但是较博来霉素组降低（P＜0.01）.各组总体样本数据行相关分析提示IL-6和IL-17表达水平具有正相关性,IL-17与TGF-β表达水平也具有正相关性.雷帕霉素干预组CD4＋CD25＋Foxp3＋细胞占CD4＋CD25＋细胞、CD4＋细胞和淋巴细胞百分比较对照组升高,CD8＋CD25＋Foxp3＋细胞的相关比例也较对照组和博来霉素组升高,并具有统计学差异（P＜0.05）.博来霉素组CD8＋CD25＋Foxp3＋细胞占CD8＋CD25＋细胞百分比较对照组高,并具有统计学差异（P＜0.01）.结论 雷帕霉素具有抑制肺纤维化的作用,其抗纤维化的作用可能通过促进调节性T细胞作者：雨林木风的诱导增殖及其对Th17细胞等效应性细胞的抑制作用发挥来实现.     

长春西汀对糖尿病大鼠静脉桥移植术后再狭窄的影响

目的探讨长春西汀(Vinp)对糖尿病静脉桥移植术后再狭窄的影响及可能作用机制。方法 36只SD大鼠随机分为对照组和Vinp组,通过建立糖尿病模型进行自体颈外静脉-颈总动脉移植,分别以生理盐水或者Vinp进行腹腔注射,于术后0周、2周及4周取移植静脉桥制作病理切片,测量内膜厚度,免疫组化法检测增殖细胞核抗原(PCNA)蛋白表达,通过细胞增殖指数定量描述。Western blot检测核转录因子(NF)-κB的磷酸化水平。结果 Vinp干预后明显减少糖尿病血管桥内膜增厚[术后2周:(17.06±5.10)μm vs.(39.79±7.84)μm,P<0.01;4周:(30.94±5.18)μm vs.(63.67±18.09)μm,P<0.01],同时还降低糖尿病静脉桥PCNA蛋白的表达[术后2周:(21.07±1.38)%vs.(28.13±1.35)%,P<0.01;4周:(31.73±1.38)%vs.(63.67±18.09)%,P<0.01],并抑制NF-κB的磷酸化(术后2周:1.08±0.42 vs.0.84±0.12,P<0.01)。结论 Vinp能有效抑制糖尿病静脉桥术后内膜增厚,其机制可能与抑制NF-κB通路,从而抑制炎症反应有关。     

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.