人附红细胞体感染FMMU白化豚鼠的研究

用附红细胞体分别感染FMMU白化豚鼠和普通花色豚鼠,同时测定两组豚鼠的红细胞免疫功能,探讨FMMU白化豚鼠的免疫学特性与病原体敏感性之间的关系。结果表明,FMMU白化豚鼠对人附红细胞体比普通花色豚鼠敏感。封闭群FMMU白化豚鼠有独特的免疫学特性,红细胞免疫功能低于普通花色豚鼠,对病原体敏感性高于普通花色豚鼠,更适于建立感染性疾病动物模型。     

SHIV病毒在猴体内的复制与传代

目的为建立SHIV艾滋病动物模型提供毒力较强的病毒株,将新合成的SHIV XJ02170病毒适应猴体,并增强其毒力。方法实验前采集猴血清并进行血清学检查和PCR检测。选出13只无SIV,STLV1,SRV D和B病毒感染的猴。第一批实验,将SHIV前病毒DNA质粒经肌肉注射到猴体内,每只500μg;SHIV病毒液,经静脉注射到猴体内,每只2ml。病毒质粒和病毒液各接种2只猴。当第一批猴体检出病毒后,10ml感染猴的全血,抗凝后静脉注射到第二批猴体内,当第二批猴检出病毒后再将10ml感染猴的全血静脉注射到第三批猴体内,连续传代4次。每批实验均定期采集血液标本,分别用肝素和EDTA抗凝,进行病毒分离;病毒基因PCR检测;CD4,CD8测定;病毒抗体检测。结果SHIV XJ02170病毒和SHIV XJ02170前病毒DNA质粒在猴体内的传代中均能分离出病毒;从传代猴的血浆和外周血单核细胞(PBMC)中检出了病毒DNA和RNA基因;CD4,CD8测定结果显示有暂时性倒置现象,后变为正常倒置与正常交替出现;在传代的猴血清中检测出特异性HIV病毒抗体。结论SHIV XJ02170病毒与SHIV XJ02170前病毒DNA质粒,均能在恒河猴体内复制。     

液血分流吸引器在剖宫产术中失血量估算的临床应用

目的为了及时诊断产后出血和正确测量剖宫产术中的失血量,及时补充血容量,降低孕产妇死亡率。方法同一剖宫术孕妇分别采用精测法（称重法和液血分流吸引器相结合）和目测法两种方法进行失血量的比较。结果目测法估算失血量明显低于精测法,结果显示用精测法对剖宫产术中进行失血量的估算是目测法的1．5倍（1．5：1）。结论在剖宫产术中精确测量失血量,及时诊断产后出血,有利于及时补充血容量,提高孕产妇身心健康。     

FMMU白化豚鼠线粒体DNA RFLP分析研究

目的研究FMMU白化豚鼠的mtDNA,并与花色豚鼠mtDNA进行多态性分析比较,以确定其独特的生物学特性是否与mtDNA相关。方法用碱变性法提取FMMU白化豚鼠以及花色豚鼠的mtDNA,并用AvaⅠ、BalⅠ等12种限制性内切酶进行酶切和限制性片段长度多态性分析。结果与结论FMMU白化豚鼠mtDNA和花色豚鼠mtDNA的相对分子质量相同,约为16.7×103;FMMU白化豚鼠与花色豚鼠两品系的mtDNA经AvaⅠ、BalⅠ等内切酶酶切后有3-8个酶切位点,酶切图谱完全相同,经RFLP分析FMMU白化豚鼠与花色豚鼠的mtDNA之间缺乏多态性。本实验没有发现FMMU白化豚鼠的独特的生物学特性与mtDNA相关。     

山西马身猪及其杂交后代H-FABP基因的PCR-RFLP检测

运用PCR-RFLP法检测马身猪及其杂交后代H-FABP基因多态性。结果表明,马身猪在Hinf Ⅰ-RFLP位点上有HH和Hh两种基因型,在HaeⅢ-RFLP位点上只有DD基因型,在HinfⅠ*-RFLP位点上有BB和Bb两种基因型,等位基因以h、D、B占优势,其频率分别为0．958、1．000和0．968。其杂交后代在HaeⅢ-RFLP位点上只检测到两种基因型DD和Dd。     

北京血站献血员戊型肝炎流行病学调查

为了解献血员戊型肝炎感染情况,对2002年7～8月向北京市血液中心义务献血的所有人员进行整群抽样并抽血,应用ELISA检测戊型肝炎病毒(HEV)IgG的感染率.结果发现:北京献血员HEV IgG总感染率为26.59%,性别、年龄、省份分布存在着差别,男性比女性感染率高,年龄越大感染率越高,来自高感染省份者感染率也高,但ALT与HEV IgG感染无关.因此,男性、年龄大、来自高感染省份具有较高的HEV感染风险,进一步研究献血员的亚临床感染对HEV的输血安全性具有重要意义.     

Spiramine N-6,一种新的抗血小板和抗血小板-中性粒细胞相互作用的物质

Spiramine N-6属粉花秀线菊植物中提取分离的二十碳二萜生物碱。本实验采用Born,Shen和Hamburger等方法分别观察了spiramine N-6在体外和体内对兔血小板聚集功能的影响。应用荧光分光光度法测定其对血小板5-羟色胺释放反应的作用,同时评价spiramine N-6对激活的血小板与中性粒细胞之间粘附反应的影响。结果表明：spiramine N-6在体外选择性抑制血小板活化因子(PAF)诱导的血小板聚集,并呈量效关系,其IC50=26μmol／L,对花生四烯酸(AA)或腺苷二磷酸(ADP)引起的血小板聚集无明显作用;spiramine N-6静注后明显抑制PAF、AA和ADP诱导的血小板聚集。Spiramine N-6呈浓度依赖性减少AA和PAF引起血小板5-羟色胺的释放,其IC50分别为64．7和33．5μmol／L。Spiramine N-6明显阻抑激活的血小板与中性粒细胞间的粘附率,其IC50为78．6μmol／L。结果提示spiramine N-6作为二十碳二萜生物碱具有较强的抗血小板和阻抑血小板一中性粒细胞相互作用的生物活性。     

Selenium and glutathione peroxidases in blood of patients with different stages of chronic renal failure

In patients with chronic renal failure (CRF) Se concentration in blood components is usually lower as compared with healthy controls. One of the five known forms of Se-dependent glutathione peroxidases (GSH-Px), the plasma GSH-Px, is synthesized primarily in the kidney. In CRF patients, plasma GSH-Px activity is reduced and the reduction increases with the progress of the disease.

The Se concentration in blood components was measured spectrofluorometrically with 2,3-diaminonaphthalene as complexing reagent. Activities of GSH-Px in red cells and in plasma were assayed by the coupled method with t-butyl hydroperoxide as substrate. The study group consisted of 150 patients in different stages of CRF. The results were compared with the values for 30 healthy subjects.

Se concentrations in whole blood and plasma of the entire group of patients were significantly lower (p < 0.01) as compared with the healthy subjects. In the incipient stage, however, the Se levels in all blood components were non-significantly lower. In whole blood and plasma the Se levels gradually decreased, reaching in the end stage values that were lower by 29 to 32% (p < 0.0001) as compared with the control group. Total protein and albumin levels in plasma of patients were significantly lower (p < 0.0001) as compared with healthy subjects and they decreased linearly with the progress of the disease. Positive and highly significant correlations were noted between total plasma protein and plasma Se concentrations (p < 0.0001) as well as between plasma albumin and plasma Se concentrations (p < 0.0001).

Red cell GSH-Px activity in the entire group of patients was lower (p < 0.05) than in the control group and did not change significantly with the progress of the disease. In plasma, however, GSH-Px activity of the entire group was lower by 33% (p < 0.0001) as compared with healthy subjects and decreased gradually with increasing renal failure. Highly significant, inverse correlations were seen between creatinine levels and plasma GSH-Px activities (p < 0.0001) as well as between urea nitrogen levels and plasma GSH-Px activities (p < 0.0001) when all stages of the disease were included.

In conclusion, patients with CRF exhibit lower Se levels in blood components as compared with healthy subjects. In whole blood and plasma these levels decrease with the progress of the disease. Plasma GSH-Px activity in patients was extremely reduced and it dramatically decreased with the progress of the illness.     

白眉蝮蛇去整合素Adinbitor的基因克隆、表达及其部分生物学活性

为了寻找去整合素家族的新成员 ,研究其在抗血栓与抗肿瘤等方面的作用 ,从中国白眉蝮蛇毒腺中提取总RNA进行RT PCR扩增 ,获得了 2 19bp的白眉蝮蛇去整合素基因 .测序结果显示 ,其与韩国的同为蝮蛇去整合素的saxatilin的DNA序列同源性为 95 8% ,蛋白质序列同源性为 91 8% ,且蛋白质中含有去整合素的特征模体 RGD .将去整合素基因进行克隆、转化与诱导后 ,得到了该蛋白的可溶性高效表达 .经组氨酸亲和层析纯化 ,获得了分子量为 9kD的均质蛋白 ,并将其命名为adinbitor .活性测定结果显示 ,adinbitor能明显抑制由碱性成纤维细胞生长因子诱导的人脐静脉血管内皮细胞ECV30 4的增殖 ,诱导ECV30 4细胞发生凋亡 ,并且呈剂量依赖性方式抑制ADP诱导的人血小板聚集     

脱氧雪腐镰刀菌烯醇对人外周血单个核细胞 HLA-I分子表达的影响

研究探讨了脱氧雪腐镰刀菌烯醇(DON)对人外周血单个核细胞HLA-I(human leucocyte cyte antigen I)分子表达影响.采用流式细胞术(FCM)和免疫印迹方法研究了不同剂量DON对体外培养人外周血单个核细胞表面HLA-I分子表达的影响及其量效关系.FCM定量检测结果表明,不同浓度DON处理均可一定程度降低人外周血单个核细胞表面HLA-I分子的表达,DON 50ng/mL、100ng/mL、1000 ng/mL和2000 ng/mL组HLA-I类分子的平均表达量分别为6.92±0.68、6.64±0.69、5.95±0.48和5.48±0.77,在50～2000ng/mL范围内随着DON浓度增加,外周血单个核细胞HLA-I分子表达降低,两者呈显著负相关(r=0.737,P＜0.01).Western印迹结果显示,大剂量DON(1000ng/mL和2000ng/mL)组人外周血单个核细胞HLA-I分子表达明显减弱.研究结果表明脱氧雪腐镰刀菌烯醇可剂量依赖地抑制体外培养的人外周血单个核细胞HLA-I分子的表达.