内皮细胞固有型一氧化氮合成酶基因G894T变异与糖尿病肾病的关系

目的：观察内皮细胞固有型一氧化氮合成酶（ecNOS）基因外显子7的Glu298Asp基因多态性与2型糖尿病肾病（DN）的关系。方法：利用聚合酶链反应限制性片段长度多态性（PCR-RFLP）技术检测了天津地区299例2型糖尿病患者和100例正常对照者的ecNOS基因外显子7的基因型，比较各组间的等位基因频率与基因型频率。结果：（1）2型糖尿病合并持续性微量白蛋白尿组（DN1组）的GT基因型和T等位基因频率高于糖尿病非肾病组（DM组）和正常对照组（C组），差异有统计学意义（P〈0．05）。（2）在2型糖尿病患者中GT基因型者较GG基因型者的DN患病率高（X^2=5．252，P=0.022）。（3）高血压、血纤维蛋白原（FIB）、ecNOS外显子7的GT基因型是2型糖尿病合并DN的独立危险因素。（4）2型糖尿病合并高血压组（DMEH组）、糖尿病非高血压组（DM1组）和C组间ecNOS基因外显子7的基因型分布和等位基因频率差异无统计学意义（P〉0．05）。结论：ecNOS基因外显子7的基因G894T（Glu298Asp）的变异可能是天津地区2型糖尿病患者DN易感性的基因标志之一，可能参与DN的启动；该基因多态性不是通过促进高血压的发生而增加DN的易感性。     

过氧化物酶体增殖体激活型受体γC161-T变异与高血压病及高血压性左心室肥厚的相关性研究

目的探讨过氧化物酶体增殖体激活型受体γ（PPARγ）C161-T变异与高血压病及高血压性左心室肥厚（LVH）的关系。方法160例高血压病患者为高血压组，116例非高血压病者为对照组，其中高血压组包括81例高血压性左心室肥厚的患者（LVH组）、79例高血压无左心室肥厚患者（无LVH组）。采用聚合酶链式反应-限制性长度多态性方法测定PPARγ基因多态性。结果高血压组与对照组PPARγ基因型的分布频率符合Hardy—Weinberg平衡，高血压组PPARγC T基因型频率显著低于对照组（21．3％对36．2％，P〈0．05），LVH组PPARγC T基因型频率显著低于无LVH组（13．6％对29．1％，P〈0．05）。结论PPARγC T基因型的多态性与高血压病及高血压性左心室肥厚相关。     

E-选择素基因多态性与新疆哈萨克族原发性高血压的相关性研究

目的探讨E-选择素（E-selectin）基因第2外显子＋G98T和第4外显子＋A561C位点单核苷酸多态性在新疆哈萨克族人群中的分布,并分析E-selectin基因多态性与原发性高血压的相关性。方法采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）的分析方法,检测了新疆哈萨克族人群无血缘关系的150例原发性高血压患者和150名对照者E-selectin基因＋G98T位点及＋A561C位点多态性,同时用生化分析仪检测相关生化指标。结果①新疆哈萨克族人群中E-selectin基因＋G98T多态性位点存在GG、GT二种基因型,其中以GG基因型发生频率最高（91．3％）,GT基因型频率次之（8．7％）;同时也存在AA、AC二种基因型,其中以AA基因型发生频率最高（88．0％）,AC基因型频率次之（12．0％）;②E-selectinAc基因型在高血压组和对照组间的分布差异存在显著性（X2=5．31,P〈0．025）。结论新疆哈萨克族人群中存在E-selectin＋G98T和＋A561C两位点单核苷酸多态性,其中A561C基因多态性与原发性高血压具有相关性,C等位基因可能是新疆哈萨克族原发性高血压发病的遗传易感基因。     

转化生长因子β1基因T869C多态性与类风湿关节炎的相关性

目的 研究转化生长因子β1（TGFβ1）基因T869C多态性与江苏地区汉族人群类风湿关节炎（RA）的相关性。方法 76例随访2年的RA患者及100例健康对照组,PCR-RFLP方法检测TG即1基因T869C多态性,比较两组间基因型及等位基因频率,并分析RA组不同基因型间临床及实验室指标的差异。结果 RA组CC基因型频率较对照组显著下降,而CT及TT基因型频率则显著升高（P〈0．05）,与对照组相比,RA组C等位基因频率下降,而T等位基因频率升高（P〈0．01）,与CC基因型相比,CT、TT基因型的RA危险度分别为2．62倍（P〈0．05）和3．60倍（P〈0．01）;RA组携带T等位基因的基因型（CT＋TT）发病年龄显著低于、CC基因型（P〈0．05）,在X线≥Ⅲ期改变比例也显著高于CC基因型（P〈0．01）。结论 TGFβ1基因多态性与RA相关,T等位基因是RA的遗传易患因子,并且与RA起病及严重程度呈正相关。     

ACE多态性与原发性高血压合并冠心病相关性研究

目的评估血管紧张素转换酶（ACE）基因多态性与原发性高血压合并冠心病的关系。方法选择高血压组64例，冠心病组81例，原发性高血压合并冠心病组92例，健康对照组122例，采用聚合酶链反应技术检测ACE基因多态性，并比较其基因型及等位基因频率。结果高血压与冠心病组ACE基因多态性分别与对照组比较均无统计学差异，原发性高血压合并冠心病组ACE基因型频率与对照组比较有统计学差异，ACE基因多态性与原发性高血压合并冠心病相关，P〈0．001。结论ACE基因多态性可能是佛山地区原发性高血压合并冠心病的危险因素。     

肿瘤坏死因子TNFα-1031T／C基因多态性与阿尔茨海默病的关系

目的 探讨肿瘤坏死因子TNFα-1031 T/C基因多态性与阿尔茨海默病（Alzheimer disease,AD）的关系.方法 采用聚合酶链式反应-限制性片段长度多态性（CR-RFLP）技术检测66例AD患者（AD组）及119名正常人（正常对照组）的TNFα-1031 T/C基因多态性.结果 AD组TNFα-1031 T/T、T/C、C/C表型频率分别为：68.18、28.78、3.03,对照组分别为57.99、41.17、0.84;AD组TNFα-1031 T、TNFα-1031 C基因频率分别为：82.58、17.42,对照组分别为：78.57、21.43;AD组TNFα-1031各种基因型频率及T、C等位基因的分布与正常对照组比较没有显著性差异（P＞0.05）.结论 TNFα-1031 T/C基因多态性与AD无明显相关性.     

妊娠期高血压疾病与血管紧张素转换酶基因和血管紧张素Ⅱ-1型受体基因多态性的相关性研究

目的探讨妊娠期高血压疾病与血管紧张素转换酶（ACE）基因插入／缺失（I／D）、血管紧张素Ⅱ-1型受体（AT1R）基因A^1166-C多态性的相关关系。方法采用聚合酶链反应．限制性内切酶片段长度多态性技术（PCR-RFLP）分别检测87例妊娠期高血压疾病患者和175例正常对照的ACE基因I／D多态性、AT1R基因A^1166-C突变位点的基因型。结果妊娠期高血压疾病患者中ACEI／D基因的ID和DD基因型频率，明显高于正常对照组，分别为41．4％和28．7％；相对于Ⅱ基因型，携带ID和DD基因型人群的OR值分别为1．981和2．347；D等位基因频率也显著高于正常对照组（49．42％），差异有显著性；相对于Ⅰ等位基因，D等位基因OR值为1．737；妊娠期高血压疾病患者中AT1R基因的AC基因型频率为33．3％；相对于AA基因型，携带AC基因型人群的OR值为1．803，C等位基因频率为17．2％，相对于A等位基因，C等位基因OR值为1．711；两组ACE和AT1R联合基因分析：相对于Ⅱ—AA联合基因型，同时携带AC-ID联合基因型的OR值为3．655；AC-DD的OR值为3．045，其余联合基因型差异均无统计学意义（P〉0．05）。结论ACE基因中D等位基因及AT，R基因A1166点突变等位基因C可能增加妊娠期高血压疾病的遗传易感性；在妊娠期高血压疾病的发生中，ACE基因及AT1R基因，可能共同对妊娠期高血压疾病的发生起作用。     

SUR1基因多态性与2型糖尿病的相关性研究

目的：探讨2型糖尿病（T2DM）与磺脲类药物受体1（SUR1）基因多态性的关系。方法：随机抽取正常人136例（正常组）及2型DM患者173例（T2DM组）外周血白细胞基因组DNA，PCR扩增SUR1基因31号外显子多态性位点的区域．用限制性酶切片段长度多态性分析其基因型，并用测序证实不同酶切图形基因型的准确性。结果：T2DM组和正常组SUR1基因31号外显子基因型及等位基因分布差别有统计学意义（P〈0．05）。结论：SUR1基因可能是中国天津地区2型糖尿病发生的易感基因。     

Graves病白细胞减少与CTLA-4基因多态性的相关性

目的：探讨天津地区汉族人Graves病（GD）合并白细胞减少，与细胞毒性T淋巴细胞相关抗原-H4（CTLA-4）基因外显子1第49位点A／G和启动子-318位点C／T二态性的相关性。方法：运用PCR—RFLP技术分析40例GD白细胞减少患者、56例GD白细胞正常患者及60例正常人CTLA-4基因、外显子1第49位点和启动子-318位点基因型．计算并比较各组基因型和等位基因频率。结果：GD组第1外显子第49位点基因型GG和等位基因G的频率明显高于正常对照组，3组人群的CTLA-4基因启动子-318位点的基因型与等位基因的分布差别无统计学意义。GD白细胞减少组与GD白细胞正常组之间基因型、等位基因的分布差别无统计学意义。结论：CTLA-4基因外显子1G49可能是天津地区汉族人Graves病的易感基因，而其与启动子-318位点可能不是天津地区汉族人GD合并白细胞减少的易感基因。     

胱硫醚β合成酶基因T833C多态性与新疆哈、汉民族原发性高血压的相关性研究

目的：探讨血浆同型半胱氨酸（Hcy）及胱硫醚β合成酶（CBS）基因T833C多态性与新疆哈萨克族（哈族）、汉民族原发性高血压（EH）的相关性。方法选取哈族239例EH患者（哈族EH组）和206例血压正常者（哈族对照组）,汉族256例EH患者（汉族EH组）和224例血压正常者（汉族对照组）为研究对象,采用全自动生化分析仪检测4组受试对象血浆中Hcy水平及相关生化指标,采用扩增阻碍突变系统（ARMS）法检测4组受试者CBS基因T833C多态性,分析Hcy、CBS基因T833C多态性与EH的相关性。结果新疆哈族、汉族EH患者血浆Hcy水平均显著高于正常对照组（P<0.05）,哈族EH组C等位基因频率明显高于哈族对照组（P<0.05）,且哈族、汉族TT基因型个体血浆Hcy水平显著低于TC基因型个体（P<0.05）;汉族EH组及汉族对照组T、C等位基因频率及TT、TC基因型频率差异均无统计学意义。结论 CBS基因T833C多态性与新疆哈族EH的发生存在相关性,与汉族EH的发生可能无相关性,其机制可能与CBS基因突变改变了Hcy的代谢有关。     

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.     

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.     

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.