人类白细胞介素10基因-592C/A多态性与SLE相关性研究

目的系统性红斑狼疮(SLE)是一种复杂的与遗传密切相关的免疫炎症性自身免疫疾病。而人类白细胞介素10(human interleukin-10,IL-10)被认为是体内最重要的抑炎细胞因子,其对炎性反应的调控起非常重要的作用,IL-10在SLE患者体内显著升高。本研究主要针对IL-10基因-592C/A多态性与SLE发病的关联性进行分析。方法抽取145例SLE患者和80例健康对照者全血标本,提取患者DNA,采用PCR和酶切、电泳的方法得到IL-10基因-592C/A位点多态性分布频率。结果 SLE患者IL-10基因-592C/A位点基因型分布和基因型频率分布与健康对照者差异无统计学意义(P>0.05)。结论 IL-10基因-592C/A多态性与SLE易感性无关。     

IL-10 基因多态性及血清水平与颅内动脉瘤发病的关系

目的 分析白细胞介素（IL）-10 基因启动子区－ 1082G/A、－ 819C/T 单核苷酸多态性（SNP）及血清 IL-10 水平与颅内动脉瘤（IAs）发病的关系。 方法 运用 PCR 及 DNA 直接测序的方法, 检测 206 例 IAs 患者（IAs 组）和 187 例非 IAs 患者（对照组）的 IL-10 基因启动子区 SNP 位点, － 1082G/A、－ 819C/T 的基因型频率和等位基因频率, χ2检验分析 IAs 组和对照组之间的差异; 采用 ELISA 法检测血清中 IL-10 水平, t 检验分析 2 组之间的差异。 结果 IL-10 基因启动子区－ 1082G/A 位点的 GG 基因型和 GA＋ AA 基因型, 以及 G 等位基因和 A 等位基因频率比较, IAs 组较对照组 GA＋ AA 基因型以及 A 等位基因频率更高, 差异均有统计学意义（P < 0.01）, GA＋ AA 基因型（OR 值为 4.137, 95%CI 2.476~6.914）和 A 等位基因（OR 值为 3.368, 95%CI 2.476~4.583）携带者有更高的 IAs 发病风险; IL-10 基因启动子区－ 819C/T 位点的 CC 基因型和 CT＋ TT 基因型以及 C 等位基因和 T 等位基因频率比较, IAs 组较对照组 CT＋ TT 基因型以及 T 等位基因频率更高, 差异均有统计学意义（P < 0.01）, CT＋ TT 基因型（OR 值为 3.393, 95% CI 1.952~5.900）和 T 等位基因（OR 值为 3.764, 95%CI 2.730~5.192）携带者有更高的 IAs 发病风险。 IAs 组血清 IL- 10 水平低于对照组（P < 0.01）。 结论 IL-10 基因启动子区 SNP 影响 IL-10 表达,IL-10 基因启动子区－ 1082G/A、－ 819C/T 多态性与 IAs 的发病有关。     

天津地区汉族人群炎症细胞因子基因多态性与冠脉支架术后再狭窄的关系

目的:探讨天津地区汉族人群白细胞介素(IL)-6-572C/G、IL-1β-511C/T及IL-10-592C/A的基因多态性及其对经皮冠状动脉介入治疗(PCI)术后再狭窄的影响。方法:437例接受PCI并进行冠脉造影随访的患者,按冠脉造影结果分为再狭窄组(166例)和非再狭窄组(271例),用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测IL-6启动子(-572)、IL-1β启动子(-511)及IL-10启动子(-592)位点基因型和等位基因频率的分布。结果:(1)2组IL-6-572C/G、IL-1β-511C/T及IL-10-592C/A基因型和等位基因频率间差异无统计学意义(均P>0.05)。(2)多因素Logistic回归分析显示脂蛋白a、急性冠状动脉综合征(ACS)、术前狭窄程度、靶病变长度为支架内再狭窄(ISR)的可能危险因素(均P<0.05),参照血管直径、支架直径为ISR的可能保护因素(均P<0.05)。结论:IL-6-572C/G、IL-1β-511C/T及IL-10-592C/A基因多态性与天津地区汉族人群再狭窄的发病无关。     

不稳定型心绞痛患者白介素-10基因1082G/A多态性分析

目的：探讨中国常州地区人群白介素-10（IL-10）基因1082G/A多态性与不稳定型心绞痛(UAP)发病的关系。方法：招募了299例研究对象并分成UAP组（n=146）和对照组（n=153）,经测序鉴定IL-10基因1082G/A多态性,并检测血糖和血脂等生化指标。结果：IL-10基因1082G/A在UAP组和对照组中均存在GG、AG和AA这3种基因型;3种基因型分布频率、等位基因分布频率与对照组相比均无统计学差异;调整对多个危险因素行多元logistic回归分析,示IL-10基因1082G/A 3种基因型与UAP的发病仍无相关性。结论：IL-10基因1082G/A多态性与中国常州地区人群UAP的发病无相关性。     

白细胞介素-10基因多态性与子宫内膜异位症的相关性

目的探讨白细胞介素-10（Interleukin-10,IL-10）基因多态性与子宫内膜异位症（Endometriosis EMs）发病的相关性。方法107例经手术治疗后,病理确认为EMs的患者,术中留取新鲜组织,对照组80例,取全血,提取DNA,采用聚合酶链式反应-限制性片段长度多态性（Polymerase chain reaction-restriction fragment length polymorphism PCR-RFLP）的方法,检测IL-10启动子区域-1082、-819、-592位点多态性。结果EMs组与对照组的IL-10启动子区域-1082、-819、-592基因型分布差异均无统计学意义（P〉0.05）。结论IL-10基因多态性与EMs的发病无明显相关性,未能够证实IL-10等位基因与EMs的遗传易感性有关。     

IL-18基因启动子-607C/A位点多态性与川崎病的关系研究1

目的：探讨IL-18基因启动子-607C／A位点多态性与川崎病（KD）的关系。方法：收集50例KD患儿（KD组）和50例健康儿童（正常组）资料,采集血液标本,分离血浆和有核细胞,用SSP-PCR法检测IL-18基因-607位点基因型、酶联免疫吸附法测定血浆IL-18含量以及免疫比浊法测定血浆CRP含量。结果：KD组患儿-607C／A位CC基因型频率为42．0％,高于正常组儿童（Jp〈0．05）;CC基因型KD患儿血浆IL-18含量显著高于CA基因型患者（P〈0．01）,前者血浆CRP含量也高于后者（P〈0．05）。结论：IL-18基因启动子-607C／A位点多态性参与KD发病,IL-18基因-607位点基因型的检测可用于预测KD发病和预后判断。     

冠心病患者MBL基因启动子区多态性

目的研究冠心病患者甘露聚糖结合凝集素(MBL)基因启动子区-550和-221位点的多态性,探讨冠心病的可能发病机制。方法提取72例冠心病患者(A组)和57例健康对照人群(C组)外周血基因DNA,采用限制性片段长度多态性分析(PCR-RFLP)法检测MBL基因启动子区-550(H/L)和-221(X/Y)位点的多态性,并行非变性的聚丙烯酰胺凝胶电泳对这2个位点的多态性进行分析。结果 A组MBL基因启动子区-550位点基因型H/L、H/H和L/L分别占2.78%、95.83%和1.39%;C组未检测出H/L和L/L基因型,H/H基因型频率为100%;A组和C组在-550位点H/L、H/H和L/L基因型频率比较差异均无统计学意义(P>0.05)。结论 MBL基因启动子区-221位点多态性可能与冠心病的发生、发展有关;而-550位点的多态性与冠心病的发生、发展可能不存在相关性。     

基质金属蛋白酶基因多态性与冠心病的关系

目的：探讨基质金属蛋白酶基因MMP-3、MMP-9和MMP-12启动子基因多态性与冠心病及其血浆水平的关系.方法：对经冠状动脉（冠脉）造影证实的急性心肌梗死患者59例（AMI组）、稳定性心绞痛患者58例（SAP组）和99例经冠脉造影证实无冠状病变的对照组进行了研究.ELISA法检测血浆MMPs的水平,多聚酶链反应-限制性内切酶片段长度多态性法（PCR-RFLP法）分析MMP-3、MMP-9、MMP-12启动子的3个部位基因型变异状况和等位基因分布频率.结果：AMI组和SAP组血浆MMP-9水平明显高于对照组（P＜0.05）.各组基因型和等位基因频率分布差异无统计学意义（P＞0.05）.未检出MMP-12启动子的-82A/G变异的基因型.不同基因型之间血浆MMPs的水平差异无统计学意义（P＞0.05）.结论：MMP-3和MMP-9启动子区域两个位点的基因变异不影响其血浆水平的表达.未发现天津地区MMP-12的-82位A/G变异的基因多态现象,血浆MMP-9水平升高与冠心病发病有关.     

单胺氧化酶A基因多态性与妊娠期高血压疾病的关系

目的探讨单胺氧化酶A（MAO-A）基因EcoRV酶切位点的基因多态性在妊娠期高血压疾病发病机制中的作用。方法采用聚合酶链式反应限制性片段长度多态性（PCR—RFLP）技术检测100例HDCP患者（病例组）及90例正常孕妇（对照组）MAO—A基因EcoRV酶切位点的基因型,并对该多态性进行统计分析。结果单胺氧化酶A基因型C／C、C／T、T／T各基因型的分布频率和C、T等位基因的分布频率在病例组与对照组之间的分布均无统计学意义（P＞0．05）。结论说明MAO—A基因EcoRV（C／T）酶切位点的基因多态性可能与HDCP的发病及病情轻重无相关性。     

天津地区胃食管反流病患者IL-1β基因多态性的研究

目的：研究白细胞介素1β（IL-1β）基因多态性与胃食管反流病（GERD）发病的相关性。方法：自134例GELID患者与182例慢性胃炎患者的胃黏膜组织提取基因组DNA，采用聚合酶链反应（PCR）方法直接测定白细胞介素1受体拮抗剂基因（IL-1RN）基因型；聚合酶链反应-限制性片段长度多态性（PCR—RFLP）方法测定IL—1β-31、Ⅱ，-1β-511基因型。通过快速尿素酶法、^14C呼气实验及组织切片染色法检测幽门螺杆菌（Hp）感染。结果：Hp阳性率在GERD组中为32．8％，慢性胃炎对照组中为62．6％，两组差异有统计学意义（P〈0．01）。IL-1RN有3种基因型，为L／L、112、2／2型，其出现频率在GERD组为78．0％、14．3％、7．7％，在对照组为55．4％、25．5％、19．1％。IL-1RN＊2基因型较L／L型发生GERD的风险下降了近70％，OR=0．29（95％CI，0．1～0．84）。IL-1β-31位点3种基因型为T／T、C/T、C／C型，在GERD组中的频率为53．7％、29．9％、16．4％，在对照组的频率为17．6％、26．4％、56％，与T／T型比较，携带C／C基因型者GERD发生的风险下降了近90％，OR=0．1（95％CI，0．04～0．23）。IL-1β-511位点3种基因型T／T,C/T，C／C，各基因型在GERD组和对照组中分布差异无统计学意义。结论：GERD患者中Hp感染率低；IL-1β—31位点和IL-1RN基因多态性与GERD发生有关，IL-1RN＊2和IL-1β—31C／C纯合子基因型是GERD的保护性因素。     

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.     

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.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

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.