获取母血中纯化胎儿有核红细胞行产前基因分析

目的　探讨获取母血中纯化胎儿细胞的有效方法。方法　对 6 0名孕龄 7～ 40周、年龄 2 1～ 30岁的孕妇外周血进行不连续密度梯度离心 ,将分离后的细胞进行制片 ,显微镜下行显微操作分离胎儿有核红细胞 ,进行 Y染色体特异性DYZ1基因的聚合酶链反应 (PCR)。结果　 (1)早孕期母血中胎儿 NRBC检出率为 2 0 % ,中孕期母血中胎儿 NRBC检出率为 71.4% ,晚孕期母血中胎儿 NRBC检出率为 6 .6 5 % ,三者之间有显著性差异 (P <0 .0 5 ) ;(2 )用显微操作技术获得的胎儿细胞数量已满足 PCR扩增所需的模板量 ,对胎儿性别进行预测 ,男胎符合率为 11/ 13(84.6 % ) ,性别符合率为 2 6 / 2 8(92 .9% )。结论　不连续密度梯度离心合并显微操作技术对于无创性早期产前基因诊断意义重大。     

Evaluating the hazard of dodecyl alkyl sulphate to natural ecosystems using indigenous protistan communities

The effect of the surfactant dodecyl alkyl sulphate (C12AS) on the structure and function of lotic protistan assemblages was examined using the Experimental Stream Facility (ESF) operated by the Procter and Gamble Company. Population- and community-level responses to C12AS were monitored on introduced substrates placed in the channels 28 days prior to dosing (mature communities) as well as those placed in the channels on day 0 of dosing (immature communities), to allow for a broad assessment of the effect of the chemical on processes contributing both to community development and maintenance. C12AS appeared to elicit a subsidy response from the native protistan assemblage which may have resulted from both positive and negative responses at trophic levels above and below the assemblage. Protistan responses to the surfactant tended to occur more rapidly and be more sensitive than those documented for invertebrates and fish. C12AS elicited a modest response (i.e. a 20% change) from several reliable parameters including community respiration (reduced dissolved oxygen at 289 g per L), protozoan species richness (increased at 63 g per L) and protozoan community composition (increased dissimilarity at 1254 g per L). Responses to C12AS exceeded expected exposures in the real world by a factor of six or greater thereby indicating that the hazard of exposure to C12AS to stream communities is low. The results of this study support the use of mesocosms as decisive tools for evaluating the hazard posed by consumer product chemicals to natural communities and ecosystems     

Mitochondrial myopathy: correlation between oxidative defect and mitochondrial DNA deletions at single fiber level

In situ hybridization combined with immunohistochemical techniques has been applied to study patients affected by mitochondrial myopathies with large mitochondrial (mt)DNA deletions. All patients' muscle biopsies showed ragged red fibers (RRFs) and cytochrome oxidase (COX) deficiency. Two digoxygenin-labeled, polymerase chain reaction (PCR)-amplifed DNAs were used as probes. One probe was designed to hybridize only with wild-type mtDNAs, while the other recognized both wild-type and deleted mtDNAs. Concomitant immunocytochemical analysis using antibodies against subunits II, III, (encoded by mtDNA) and IV (encoded by nuclear DNA) of COX was carried out. In our patients deleted mtDNAs are overexpressed in COX-negative RRFs, while wild-type mtDNAs are decreased in the same fibers. Immunohistochemistry studies show that COX IV is overexpressed in RRFs and that COX II and COX III subunits are still present. Deleted mtDNAs are spatially segregated in muscle fibers, where they interfere with the local population of normal mitochondrial genomes, causing a regional deficiency of the mitochondrial respiratory activity.This work was supported by the Associazione Amici del Centro Dino Ferrari     

A Simple and Rapid Fluorometric Determination Method of α1-Acid Glycoprotein in Serum Using Quinaldine Red

We examined different fluorescent probes suitable for fluorometric determination of ₁-acid glycoprotein (AGP) in serum. Quinaldine red (QR) was shown to bind strongly and selectively to AGP. Taking advantage of the enhanced fluorescence of QR in the presence of AGP, we developed a direct method for the determination of serum AGP without removal of other serum proteins such as albumin. AGP concentrations in serum of healthy volunteers and patients correlated well with results from the conventional single radial immunodiffusion (SRID) method (r = 0.93, slope = 1). The newly developed method is faster and has a larger analytical concentration range than the SRID method. This method can also be used to determine AGP in serum of experimental animals, and it can serve to monitor AGP serum concentrations for pharmacokinetic evaluation of basic drugs.     

泰兴地区肝癌高发因素研究

探讨了泰兴地区人群肝癌的高发因素。采用ELISA法分别检测人群体内HBV感染水平和AFT—SHA含量及生活饮用水水体内MC含量。结果发现该地区人群HBsAg携带率为29．5％,HBV感染阳性率68.1％,肝癌高发乡镇人群HBsAg携带率和HBV感染阳性率分别高达45．8％和94．7％;人群体内AFT—HSA检出率达100％,平均含量为16．39pg／ng;高发乡镇人群体内AFT—HSA平均含量31．99pgM;生活饮用水水体内MC阳性检出率13．2％,河水、沟塘水和浅井水内MC的平均含量分别为36ng／L,29ng／L和25ng／L．结论：人群中的HBV高感染;黄曲霉素强暴露;生活饮用水水体内MC的普遍污染是泰兴地区肝癌高发的三个重要因素．     

锥兰联合茜素红活细胞染色法观察角膜中央碱烧伤内皮细胞愈合过程

目的:研究角膜碱烧伤内皮细胞的愈合过程。方法:锥兰联合茜素红内皮细胞染色法对角膜中央碱烧伤内皮细胞形态学变化进行观察。结果:伤后20分钟内皮细胞已破坏。72小时,损伤区的周边内皮细胞变形向创面内迁移。第7天后缺损区大部分范围均可见梭形细胞覆盖。结论:兔碱烧伤损伤区内皮细胞的修复由邻近内皮细胞变形,增殖和移行长入创面内完成,修复的内皮细胞具有纤维细胞特征。眼科学报1999;15:218-220。     

Concomitant blockade of P2X-receptors and ecto-nucleotidases by P2-receptor antagonists: functional consequences in rat vas deferens

In order to assess the consequences of a concomitant blockade of P2X-receptors and ecto-nucleotidases, effects of 13 P2-receptor antagonists were investigated on contractions of the rat vas deferens elicited by α,β-methylene ATP (α,β-MeATP) and ATP and on the removal of ATP from the incubation medium by vas deferens tissue. Increasing concentrations of all antagonists reduced and finally abolished contractions elicited by α,β-MeATP (3 μM), with IC₅₀-values ranging from 1.1 to 100 μM. Pyridoxalphosphate-6-azophenyl-2’,4’-disulphonate (PPADS), 6-azophenyl-4-amino-5-hydroxy-naphthalene-1,3-disulphonate (NH02), 4,4’-diisothiocyanatostilbene-2,2’-disulphonate (DIDS) and uniblue A also progressively reduced and finally abolished contractions elicited by ATP (1 mM). 8,8’-[Carbonylbis(imino-3,1-phenylenecarbonyl-imino)]-bis-(1,3,5-naphthalenetrisulphonate) (NF023), sura- min, pyridoxalphosphate-6-azophenyl-2’,5’-disulphonate (iso-PPADS), trypan blue and reactive blue 19, in contrast, caused only partial blockade, by 34–43% maximally; reactive blue 2 and reactive red 2 had no effect; and 6,6’-(1,1’-biphenyl-4,4’-diylbisazo)-bis-4-amino-5-hydroxy-naphtha-lene-1,3-disulphonate (NH01) and Evans blue even enhan- ced the response to ATP. For antagonists causing full or partial inhibition, the IC₅₀-values against ATP were close to those against α,β-MeATP. All antagonists attenuated the removal of ATP, with IC_25%-values ranging from 0.8 μM to >320 μM. The results confirm the frequent combination, in one antagonist molecule, of P2-receptor blockade and blockade of ecto-nucleotidases. This dual action underlies the effect of such compounds on contractions of the vas deferens elicited by ATP which, for certain substances (e.g., suramin, reactive blue 2), can be explained by a simple model in which the antagonist simultaneously blocks the degradation of ATP and a single contraction-mediating receptor (P2X₁). Several observations, however, do not conform with this model, and the existence of multiple contraction-mediating receptors for ATP or multiple, pharmacologically distinct ecto-nucleotidases has to be considered. Received: 23 October 1998 / Accepted: 11 January 1999     

消红散对实验性肾小球肾炎小鼠红细胞免疫功能作用机制的研究

目的 研究具有益气健脾摄血功效的消红散对肾小球肾炎(GN)小鼠红细胞免疫功能的影响。方法复制免疫性GN小鼠模型，检测小鼠血中C3b受体花环率及免疫复合物(CIC)、红细胞免疫黏附促进因子(RFER)、红细胞免疫黏附抑制因子(RFIR)的变化。结果消红散能增加GN小鼠的红细胞免疫功能，使C3b受体的数目增加，促使红细胞清除免疫复合物，减少CIC在肾小球基底膜沉积，缓解炎症或CIC介导的病理过程，且能使RFER活性升高，RFIR活性下降。结论 消红散对红细胞免疫功能具有双向调节作用。     

柴胡、赤芍与醋柴胡、杭白芍对大鼠高脂血症作用的实验研究

目的观察药对柴胡、赤芍与醋柴胡、杭白芍,对高脂血症大鼠脂质代谢、血小板功能影响,比较它们作用环节的差异.方法本实验采用高脂应激造成大鼠高脂血症血瘀模型方法,观察大鼠血瘀模型脂质代谢、血小板功能的变化.结果醋柴胡、杭白芍、柴胡、赤芍对TC(cholesterol)均有显著降低作用,醋柴胡、杭白芍强于柴胡、赤芍,而两者对LDL-C(low density lipoprotein)、Lp(a)[lipoprotein(a)]也有显著降低作用,但柴胡、赤芍作用较强,和模型组比较有明显差异(P＜0.05和P＜0.01);醋柴胡、杭白芍对TG (triglyceride)有显著降低作用,和模型组比较有明显差异(P<0.01); 柴胡、赤芍对血小板聚集率和活化阳性率均有显著降低作用(P<0.05).结论柴胡、赤芍与醋柴胡、杭白芍可作用于引起动脉粥样硬化的不同环节,柴胡、赤芍药对在抑制血小板聚集、降低血小板活化率方面优于醋柴胡、杭白芍.