多形核粒细胞质膜结合酶活性的氧化损伤及IH—764—3的抗损伤作用

用乙酸肉豆寇咐醇酸刺激人多形核粒细胞可导致其质膜结合酶活性下降，碱性磷酸酶、５＇－核苷酸酶和中性内肽酶活性分别降至３６．６１％、５８．５３％和３０．７９％。Ｏ２及Ｈ２Ｏ２生成抑制试验证明，ＰＭＡ刺激后ＰＭＮ刺激后ＰＭＮ质膜结合酶活性的损伤乃活性氧物质所致。ＩＨ－７６４－３是自中药提取的一种单体成分，经实验证明具有抗氧化损伤，保护ＰＭＮ质膜结合酶的作用，该抗损伤作用可能与其抑制Ｈ２Ｏ２生成有关。     

乙型肝炎患者血清抗HBs独特型抗体免疫复合物的检测

用聚乙二醇沉淀，胰蛋白酶处理乙型肝炎（乙肝）患者血清后，再用酶联免疫吸附法检测检样中的含抗ＨＢｓ独特型抗体免疫复合物（抗ＨＢｓ－Ａｂ＿２－ＩＣ）。结果发现，乙肝患者血清中ＩｇＧ、ＩｇＭ类抗ＨＢｓ－Ａｂ，－ＩＣ检出总阳性率分别为：急性乙型肝炎１４．５％（９／６２），慢性活动性肝炎２１９％（２８／１２８），慢性迁延性肝炎１２．５％（２／１６）。抗ＨＢｓ－Ａｂ＿２－ＩＣ阳性者的抗ＨＢｓ独特型抗体检出率８４．１％（３３／３９），且乙型肝炎病毒的血清学标志阳性率显著高于抗ＨＢｓ－Ａｂ＿２－ＩＣ阴性者。表明，乙肝患者体内存在抗ＨＢｓ－Ａｂ＿２－ＩＣ，抗ＨＢｓ独特型抗体（抗ＨＢｓ－Ａｂ＿２）可能通过与抗ＨＢｓ结合，削弱和影响抗ＨＢｓ中和、清除乙肝病毒抗原的作用。     

家兔脑缺血再灌注损伤机制的研究

用家兔四动脉闭塞法建立急性完全性脑缺血后给予再灌注，观察缺血前后及再灌注后脑组织部分电解质和含水量、脑组织和血液丙二醛（ＭＤＡ）、环核昔酸（ｃＡＭＰ、ｃＧＭＰ）、血栓烷Ｂ＿２（ＴｘＢ＿２）、６－酮－前列腺素Ｆ＿（１α）（６－ｋｅｔｏ－ＰＧＦ＿（１α）含量、超氧化物歧化酶（ＳＯＤ）和谷胱苷肽过氧化物酶（ＧＳＨ－Ｐｘ）活性改变。结果：再灌注后实验组脑组织Ｃａ￣（２＋）、ＭＤＡ、ＴｘＢ＿２和ｃＡＭＰ较对照组升高（Ｐ＜０．０５），ＧＳＨ－Ｐｘ活性、６－ｋｅｔｏ－ＰＧＦ＿（１α）含量降低（Ｐ＜０．０５）；实验组血液中ｃＡＭＰ含量高于对照组（Ｐ＜０．０５）。提示脑组织Ｃａ￣（２＋）过载、氧自由基增多、ｃＡＭＰ／ｃＯＭＰ和ＰＧＩ＿２／ＴＸＡ＿２比值异常在脑缺血再灌注损伤中起着重要的作用。     

Graves病甲状腺NADH-细胞色素b5还原酶活性与碘有机化关系

目的 探明ＮＡＤＨ－细胞色素ｂ５还原酶（ｂ５Ｒ）在甲状腺激素合成中的作用。方法：应用ＮＡＤＨ－高铁氰化钾等测定甲状腺ｂ５Ｒ和Ｈ２Ｏ２酶活性,并在体外增减 机化系统,观察ｂ５Ｒ活性有机碘形成关系。结果 ：Ｇｒａｖｅｓｅ病（ＧＤ）ｂ５Ｒ活性 明显增高,所形成的有机碘也明显增多ｂ５Ｒ抑制剂后有机碘形成又明显减少。ＧＤ时Ｈ２Ｏ２酶活性无改变,结果：ｂ５Ｒ在甲状腺激素合成中起重要作用。抑制ｂ５Ｒ活性就能抑制     

ACTH及电针对甲醛痛敏大鼠脊髓NOS阳性神经元增多的影响

本文采用ＮＡＤＰＨ－ｄ组化染色技术,研究鞘内注射（ｉ．ｔ．）ＡＣＴＨ及电针刺激（ＥＡ）对甲醛痛敏大鼠脊髓背角浅层ＮＯＳ阳性神经元增多的影响。结果显示ＡＣＴＨ（０．５Ｕ,ｉ．ｔ．）及电针（１ｍＡ５０Ｈｚ,５ｍＡ５Ｈｚ,１ｍＡ５Ｈｚ）刺激“夹脊穴”３０ｍｉｎ均可显著抑制脊髓背角浅层ＮＯＳ阳性神经元增多;ｉ．ｔ．ＡＣＴＨ（０．５Ｕ）和电针刺激（１ｍＡ５Ｈｚ）同时给予时,抑制作用显著增加;ｉ．ｔ．ＮＯ前体     

前列腺素E1对心肌缺血—再灌注损伤保护作用的观察

目的：研究心肌缺血再灌注时人体外周血中性粒细胞（ＰＭＮ）的数量、ＰＭＮ聚集率、内皮素（ＥＴ）、脂质过氧化物（ＬＰＯ）、超氧化物歧化酶（ＳＯＤ）及过氧化氢酶（ＣＡＴ）的变化，并观察前列腺素Ｅ１（ＰＧＥ１）对心肌缺血再灌注损伤的保护作用。方法：将３６例急性心肌梗死患者随机分为２组，每组１８例。治疗组给予ＰＧＥ１加尿激酶和硝酸甘油静滴，对照组给予尿激酶和硝酸甘油静滴，于再灌注前后分别采静脉血测定上述指标。结果：心肌缺血时外周血ＰＭＮ数量无显著增多，但ＰＭＮ活性显著增强，血ＥＴ及ＬＰＯ水平增高，血ＳＯＤ和ＣＡＴ活性下降；再灌注后，ＰＭＮ数量无明显增多（Ｐ均＞０．０５），但ＰＭＮ活性进一步显著增强，ＥＴ及ＬＰＯ水平进一步显著增高，而ＳＯＤ和ＣＡＴ活性则进一步显著下降。使用ＰＧＥ１治疗后结果则与上述情况相反。结论：ＰＭＮ及ＥＴ在心肌缺血再灌注损伤过程中起重要作用，ＰＧＥ１可以抑制ＰＭＮ活性，保护ＳＯＤ和ＣＡＴ活性，减少ＥＴ及ＬＰＯ产生，减轻过氧化反应，对心肌缺血再灌注损伤有显著的保护作用。     

费乐地平对兔心肌缺血—再灌注损伤的保护作用

目的：本研究旨在探讨心肌再灌注损伤（ＲＩ）的机制并评价费乐地平（Ｆｅｌｏｄｉｐｉｎｅ，Ｆｅｌ）对ＲＩ的保护作用及其机制。方法：用家兔心肌缺血再灌注模型，分别设对照组、单纯缺血再灌注组及再灌注＋Ｆｅｌ治疗组，设再灌注０．５、１．５和６．０小时３个时相点，测定心肌Ｃａ２＋、丙二醛（ＭＤＡ）含量、中性粒细胞（ＰＭＮ）浸润数、Ｎａ＋Ｋ＋ＡＴＰ酶及Ｃａ２＋Ｍｇ２＋ＡＴＰ酶活性，并观察心肌超微结构改变及测定心肌梗塞范围。结果：心肌再灌注后Ｃａ２＋、ＭＤＡ含量及ＰＭＮ浸润数明显增高，Ｎａ＋Ｋ＋ＡＴＰ酶和Ｃａ２＋Ｍｇ２＋ＡＴＰ酶活性明显降低，其改变以Ｃａ２＋升高发生最快；Ｆｅｌ可使上述指标改变的程度明显减轻（Ｐ＜０．０５～０．０１），并可缩小心肌梗塞范围，改善其超微结构的变化。结论：心肌ＲＩ是多种因素于不同时间所致的一种复合性损伤，Ｆｅｌ对心肌ＲＩ有保护作用，其保护机制可能与其对心肌能量依赖性酶活性的改善有关。     

酶偶联法测定红细胞谷胱甘肽过氧化物酶

根据酶偶联催化反应机理，通过二次作图法测定了过氧化物酶（Ｓ＿１）和Ｈ＿２Ｏ＿２（Ｓ＿２））的米氏常数（Ｋｍ），分别为０．０６９ｍｍｏｌ／Ｌ和０．１５ｍｍｏｌ／Ｌ。通过理论计算和实验分析确定Ｓ＿１的最适底物浓度为１．５ｍｍｏｌ／Ｌ，Ｓ－２最适底物浓度为２．２５ｍｍｏｌ／Ｌ。实验证明指示酶（Ｇ·Ｒ）用量应＞０．２５Ｕ／ｍｌ反应液，本法采用０．３９Ｕ／ｍｌ反应液并就ｐＨ、温度、Ｈｂ干扰等因素对实验影响进行了系统研究，优选了最佳条件。本法批内变异系数为３．８％，批间变异系数为５．２％。     

急性髓细胞白血病M_（2b）型AML_1－ETO融合基因转录本的检测

急性髓细胞白血病（ＡＭＬ）－Ｍ＿（２ｂ）患者约９０％有ｔ（８；２１）。已经证明它导致ＡＭＬ＿１－ＥＴＯ融合基因。我们应用逆转录酶／聚合酶链反应（ＲＴ／ＰＣＲ）检测了２２例ＡＭＬ＿１－Ｍ＿（２ｂ）其中ｔ（８；２１）阳性１６例，阴性２例，不能确定有无ｔ（８；２１）的４例，发现全部２２例患者均可检出ＡＭＬ＿１－ＥＴＯ融合基因转录本。同时在５例ｔ（８；２１）阴性的ＡＭＬ中，包括Ｍ＿（２ａ）２例，Ｍ＿４２例，Ｍ＿３１例，均未检出上述融合基因转录本。我们的初步结果提示ＡＭＬ＿１－ＥＴＯ融合基因可以作为ＡＭＬ－Ｍ＿（２ｂ）的基因标志。     

HCV感染基因型病毒复制程度与肝病关系的研究

采用限制性内切酶Ｓａｕ３Ａｌ和ＨａｅⅢ对５４例ＲＴ－ＰＣＲ法ＨＣＶＲＮＡ阳性的５＇ＮＣ区扩增产物进行酶切分型；ＰＣＲ滴定法半定量分析ＨＣＶＲＮＡ含量。结果显示４种酶切类型：ＨＣＶⅠ型感染３．７％（２／５４），ＨＣＶⅡ型感染７７．８％（４２／５４），ＨＣＶⅢ、Ⅳ型感染１１．１％（６／５４），ＨＣＶⅠ、Ⅱ／Ⅲ、Ⅳ型混合感染７．４％（４／５４），ＨＣＶⅡ型感染阳性率在ＡＨ、ＣＡＨ、ＬＣ三组间无显著差异（     

佛波醇酯(PMA)对中性粒细胞(Np)质膜结合酶的钝化作用

本文探索了佛波醇酯(PMA)激活的中性粒细胞(Np)对其质膜结合酶的影响,发现低浓度PMA激活的Np可以引起其质膜酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、5’-核苷酸酶(5’-NT)、中性内肽酶(NEP)和乙酰胆碱酯酶(AChE)5种膜结合酶的钝化,酶活性分别下降57％、61％、52％、34％和66％。NBT试验证明PMA激活的Np产生大量的活性氧物质,而且包括活性氧清除剂在内的多种防护药物可以有效地拮抗该钝化作用,如SOD+CAT合剂可使5种酶活性分别维持在87％、88％、100％、51％和94％,证明钝化作用系产生的活性氧物质所致。     

转染HSP70基因对过氧化氢所致RAW264.7细胞损伤的影响

[目的]观察HSP70基因过表达是否能减轻过氧化氢所致的RAW264.7巨噬细胞损伤.[方法]用脂质体包裹重组质粒pcDNA3.1-HSP70导入RAW264.7细胞株,经G418筛选阳性克隆,用Western blot方法检测HSP70的表达情况;通过测定细胞存活率、LDH释放率,观察了HSP70基因过表达对过氧化氢所致RAW264.7细胞损伤的影响.[结果]获得了HSP70过表达的RAW264.7细胞系;HSP70过表达组细胞的存活率及LDH释放率与对照组比有明显差异.[结论]转染HSP70基因对过氧化氢所致RAW264.7细胞损伤有明显的保护作用.     

有机物对H2O2低温等离子灭菌效果影响的研究

目的探讨有机物对H2O2等离子低温灭菌效果的影响,为进一步规范内镜消毒提供理论依据。方法用聚四氟乙烯软管模拟内镜,以大肠杆菌悬液制备染菌载体,小牛血清作为有机物,研究有机物对H2O2等离子低温灭菌效果的影响。结果无小牛血清存在的情况下,H2O2等离子低温灭菌5min后杀灭率可以达到100%,25%和50%小牛血清均对杀灭菌效果有影响,作用时间延长后杀灭率可以达到100%。结论有机物对H2O2等离子低温灭菌系统灭菌效果有显著的影响,在有机物存在时其杀菌作用明显减弱,且浓度越高影响越大,说明器械灭菌前彻底清洁尤为重要。     

Chitotriosidase activity in plasma and mononuclear and polymorphonuclear leukocyte populations

In the general population, about 5% of individuals are homozygotic and 35% are heterozygotic carriers for chitotriosidase (ChT) deficiency. Activated macrophages are considered to be the main source of plasma ChT activity, which permits the biochemical characterization of homozygote deficients. However, in the case of detecting heterozygotic carriers, the results are often inconclusive. The activities of ChT in plasma and mononuclear (MN) and polymorphonuclear (PMN) leukocytes were determined in 169 control subjects (72 males and 97 females) with a mean age (+/- SD) of 47.5+/-9.7 years (range 18-96 years). The specific enzyme activity was in PMN leukocytes >MN leukocytes >plasma, with a highly significant partial correlation being found between the activities of ChT in plasma and PMN leukocytes (r=0.578, P<0.001). A significant correlation was found between the age of the patients studied and plasma ChT activity (r=0.568, P<0.001). No significant correlation was found for enzyme activities in MN (r=0.105) or in PMN leukocytes (r=0.043). The results obtained suggest that, in normal physiological conditions, PMN leukocytes may secrete ChT to the plasma. Although the activities of ChT in MN and PMN leukocytes are not affected by demographic factors, it is not possible to use them for the biochemical detection of ChT-deficient heterozygotic carriers.     

有机物对H2O2低温等离子灭菌效果影响的研究

目的通过有机物对H2O2等离子低温灭菌效果的影响,寻找内镜消毒灭菌中的薄弱环节,为进一步规范内镜消毒提供理论依据。方法选取造成医院感染源之一的大肠艾希菌为代表,取小牛血清作为有机物,并取25%、50%的小牛血清浓度为可变因素,研究有机物对H2O2等离子低温灭菌效果的影响。结果无小牛血清存在的情况下,H2O2等离子低温灭菌系统作用5 min后杀灭率可以达到100%,而有25%和50%小牛血清存在时,对杀菌效果有影响。结论有机物对H2O2等离子低温灭菌系统灭菌效果有显著的影响,在有机物存在时其杀菌作用明显减弱,且浓度越高影响越大。因此,器械灭菌前必须彻底清洗干净。     

H_2O_2致PC12细胞凋亡与细胞内14-3-3蛋白表达水平下调的关系

目的:探讨用H2O2处理PC12细胞制作PD氧化应激细胞模型的最佳浓度与最适处理时间,以及H2O2致PC12细胞凋亡与14-3-3蛋白表达水平的关系。方法:采用MTT法、荧光分光光度计、流式细胞术及免疫印迹技术分别检测PC12细胞活力、Caspase酶活性、细胞凋亡率以及14-3-3蛋白表达量。结果:当H2O2浓度为100μmol/L时Caspase活性最高;100μmol/LH2O2处理PC12细胞24h时细胞凋亡率最高;PC12细胞的凋亡率与14-3-3蛋白表达的水平呈显著负相关。结论:100μmol/L和24h分别为用H2O2制作PD氧化应激模型的最佳浓度和处理时间,H2O2致PC12细胞的凋亡与细胞内14-3-3蛋白表达水平下调有关。     

二氧化氯协同消毒器产气量影响因素的研究

由美国引进的二氧化氯协同消毒器可产生ClO_2、O_3、H_2O_2等气体对水进行协同杀菌。本研究在实验室测定了电解电压、电解电流、电解质浓度与阳极有效面积对消毒器产气量的影响。     

ACUTE DESTRUCTION BY HUMORAL ANTIBODY OF RAT SKIN GRAFTED TO MICE : THE ROLE OF COMPLEMENT AND POLYMORPHONUCLEAR LEUKOCYTES

A study has been made of the roles played by complement and polymorphonuclear leukocytes (PMN) in the acute destruction of xenografts of rat skin that follows injection of their hosts with antisera specifically reactive with graft antigens. The rat skin was grafted onto mice whose immune responses were suppressed by removal of the thymus and a brief course of treatment with rabbit antimouse lymphocyte serum. At about 2 wk after grafting the mice were injected intravenously or intraperitoneally with mouse antirat serum (MARS). This time interval was chosen because it avoided the complications that might be associated with either the process of healing in or with incipient rejection. Signs of graft damage were evident as early as 10 min after the injection of MARS, and in most animals so injected the grafts were completely destroyed within 24–48 h. The role of complement (C) in this acute destructive process is indicated by the results of three lines of experimentation. (a) Non-C-fixing antibodies or antibody fragments failed to cause damage to the grafts. Indeed, both chicken antirat serum and F(ab')₂ fragments from rabbit antirat serum completely protected the grafts against the effects of MARS that was administered 24 h later. (b) When mice were depleted of hemolytic C by treatment with cobra venom factor or heat-aggregated gamma globulin, the damage caused by MARS was greatly reduced or completely inhibited. (c) In mice with a genetically determined absence of C5 much greater quantities of MARS were required to cause graft damage; the tempo of the destructive process was consistently slower; and a greater number of grafts recovered from the initial inflammatory process than was the case for animals with an intact complement system. The participation of PMN in serum-mediated destruction of grafts was initially suggested by the results of microscope examination of fixed tissues. The essential role of these cells in the process is indicated by the failure of MARS to cause tissue damage in mice whose circulating PMN have been reduced to very low levels by treatment with nitrogen mustard or more specifically with an anti-PMN serum. The absence of tissue damage when circulating PMN are reduced but C levels are normal suggests that C-mediated cytolysis is unimportant in graft destruction and that the role of C lies in its ability to generate chemotactic factors. The latter may then attract the PMN that provide mediators of tissue damage.     

Endotoxin-induced human and porcine leucocyte reactions in vitro

Tube migration of a fixed number of PMN cells in plasma (5 X 10(9) cells/l) was approximately reduced by 50% when 1 ml of cell suspension was exposed to 20 micrograms E. coli endotoxin in 100 microliter NaCl for 2 hours. Procoagulant activity in monocytes increased approximately eight-fold when 1 ml of whole blood was exposed to 2 ng endotoxin in 10 microliters NaCl for 2 hours. Chemiluminescence in both PMN cells (5 X 10(9) cells/l) and mononuclear cells (2 X 10(9) cells/l) in plasma was markedly increased when 100 microliters cell suspension was added to 100 microliters Luminol, exposed to 20 micrograms endotoxin in 100 microliters NaCl and tested immediately. Decreased lysosomal enzyme levels in PMN and mononuclear cells were demonstrated when 1 ml cells in plasma (the same cell numbers as aforementioned) were incubated for 4 hours at 37 degrees C with 200 micrograms endotoxin in 100 microliters NaCl. Similar results were obtained in human and porcine leucocytes, making the pig a suitable animal for studies of humoral and cellular reactions to infections complications following trauma and major surgery.     

Purified human plasma kallikrein aggregates human blood neutrophils.

Exposure of human blood polymorphonuclear leukocytes (PMN) to purified active plasma kallikrein resulted in PMN aggregation when kallikrein was present at concentrations ranging from 0.4 to 0.6 U/ml (0.18-0.27 microM). Kallikrein-induced PMN aggregation was not mediated through C5-derived peptides, because identical responses were observed whether or not kallikrein had been preincubated with an antibody to C5. Moreover, kallikrein was specific for aggregating PMN, because no aggregation was observed with Factor XII active fragments (23 nM), Factor XIa (0.6 U/ml or 15nM), thrombin (1.6 microM), plasmin (2 microM), porcine pancreatic elastase (2 microM), bovine pancreatic chymotrypsin (2 microM), or bradykinin (1 microM). Bovine pancreatic trypsin (2 microM) aggregated PMN, but to a lesser extent than kallikrein (0.18 microM). Kallikrein was a potent aggregant agent for PMN because similar responses were observed with kallikrein (0.5 U/ml or 0.23 microM) and an optimal dose (0.2 microM) of N-formyl-methionyl-leucyl-phenylalanine. In addition, PMN incubation with kallikrein resulted in stimulation of their oxidative metabolism as assessed by an increased oxygen uptake. Neutropenia and leukostasis observed in diseases associated with activation of the contact phase system may be the result of PMN aggregation by plasma kallikrein.