不对称PCR制备单链探针检测登革Ⅱ型病毒复制型RNA和复制中间体RNA

登革病毒是具包膜的单股正链RNA虫媒病毒 ,病毒的复制过程发生在感染细胞胞浆 ,复制型 (RF)RNA是病毒半保留复制的循环模板 ,复制中间体 (RI)RNA的合成则是病毒复制所必需的。经RT PCR获得的DNA模板进行不对称PCR扩增 ,当限制性引物终浓度为 2 5 0nmol L ,两引物比例为 1 0 0∶1时 ,即得到不对称PCR的预计单链和双链DNA产物。此单链产物用于标记探针进行核酸杂交。结果表明不对称PCR制备单链探针进行核酸杂交可用于检测病毒复制型RNA和复制中间体RNA的合成     

Evaluation of anti-parvovirus B19 activity in sera by assay using quantitative polymerase chain reaction

Human parvovirus B19 (B19) infects cells of erythroid lineage. Production of neutralizing antibodies (Abs) is indispensable for recovery from B19-related disease state. In this study, we used a convenient method to measure neutralizing activities in human sera by using a real-time quantitative PCR based assay. Erythroid cell line KU812Ep6 was incubated with test sera before infection with B19 virus. The copy number of B19-DNA in cultures was decreased in the presence of the sera from patients who recovered from acute B19 infection, whereas no decrease in B19-DNA was in cultures incubated with sera from healthy volunteers who had no B19 infection. The decrease in B19-DNA copy number was calculated and the inhibition percentage was expressed as neutralizing activity to B19. A clinical study showed that the levels of neutralizing ability were high in patients who recovered soon after acute B19 infection, but were low in some patients with a prolonged clinical course for recovery from B19 infection. This method is simple and convenient compared with methods described previously, showing its usefulness to evaluate the neutralizing activity to B19.     

Effects of x-ray and CT image enhancements on the robustness and accuracy of a rigid 3D/2D image registration

A rigid body three-dimensional/two-dimensional (3D/2D) registration method has been implemented using mutual information, gradient ascent, and 3D texturemap-based digitally reconstructed radiographs. Nine combinations of commonly used x-ray and computed tomography (CT) image enhancement methods, including window leveling, histogram equalization, and adaptive histogram equalization, were examined to assess their effects on accuracy and robustness of the registration method. From a set of experiments using an anthropomorphic chest phantom, we were able to draw several conclusions. First, the CT and x-ray preprocessing combination with the widest attraction range was the one that linearly stretched the histograms onto the entire display range on both CT and x-ray images. The average attraction ranges of this combination were 71.3 mm and 61.3 deg in the translation and rotation dimensions, respectively, and the average errors were 0.12 deg and 0.47 mm. Second, the combination of the CT image with tissue and bone information and the x-ray images with adaptive histogram equalization also showed subvoxel accuracy, especially the best in the translation dimensions. However, its attraction ranges were the smallest among the examined combinations (on average 36 mm and 19 deg). Last the bone-only information on the CT image did not show convergency property to the correct registration.     

A new model for inflammation-induced preterm birth: the role of platelet-activating factor and Toll-like receptor-4

Preterm birth is a leading cause of neonatal morbidity and mortality. Despite a growing body of evidence correlating inflammation with preterm birth, the signal transduction pathways responsible for the emptying of the uterus in the setting of intrauterine inflammation has not been elucidated. We now report a unique, reproducible mouse model of localized intrauterine inflammation. This model results in 100% preterm delivery with no maternal mortality. Using our model, we also show that platelet-activating factor is a crucial mediator of both inflammation-induced preterm birth and fetal demise. Using C3H/HeJ mice, we demonstrate that toll-like receptor-4 (TLR-4) plays a role in lipopolysaccharide-induced preterm birth but not in inflammation-induced fetal death. Immunohistochemistry studies demonstrate the presence of the platelet-activating factor receptor in both endometrial glands and smooth muscle in uterine tissues. Molecular studies demonstrate the differential expression of platelet-activating factor receptor and TLR-4 in uterine and cervical tissue throughout gestation. Quantitative polymerase chain reaction revealed an up-regulation of TLR-4 in the fundal region of the uterus in response to intrauterine inflammation. The use of this model will increase our understanding of the significant clinical problem of inflammation-induced preterm birth and will elucidate signal transduction pathways involved in an inflammatory state.     

再生障碍性贫血患者细胞因子水平观察

为进一步探讨细胞免疫对ＡＡ发病的影响，阐明细胞因子在ＡＡ患者中变化的基础与临床意义，采用酶联免疫试剂盒ＥＬＩＳＡ法对３８例ＡＡ患者及２０例正常人外周血单个核细胞（ＰＢＭＮＣ）培养上清诱生Ｇ－ＣＳＦ，ＩＬ－６，ＴＮＦα，ＩＦＮα及ＩＬ－８水平进行测定，同时采用改良ＡＰＡＡＰ法观察外周血Ｔ细胞亚群及ＨＬＡ－ＤＲ抗原表达，结果ＡＡ患者ＰＢＭＮＣ培养上清中Ｇ－ＣＳＦ阳性率减低，ＩＬ－６，ＴＮＦαＩＦＮα及     

散发性戊型肝炎病毒感染的诊断

用基因工程重组的戊型肝炎病毒基因结构区第二码框架和第二读码框架具有免疫表位的嵌合抗原，建立了间接酶联免疫法，检测散发性急性肝炎病人血清中抗－ＨＥＶＩｇＧ和ＩｇＭ抗体。在４６例急性肝炎病人中出抗－ＨＥＶＩｇＧ抗体阳性７例，阳性率为１５．２２％，７例ＩｇＧ抗体阳性中，有５例ＩｇＭ抗体也阳性，占７１．４％。     

Radiation-induced lung injury in vivo: Expression of transforming growth factor—Beta precedes fibrosis

Cytokine release from irradiated cells has been postulated to start soon after irradiation preceding detectable clinical and pathological manifestation of lung injury. The expression of transforming growth factor beta (TGF), a fibrogenic and radiation-inducible cytokine, was studied from 1–16 weeks after the 15 and 30 Gray (Gy) of thoracic irradiation to rats. Thoracic irradiation caused an increase in TGF protein in bronchoalveolar lavage (BAL) fluid peaking at 3–6 weeks as compared to sham-irradiated control rats. Steady state TGF mRNA expression as shown by whole lung northern blot assay paralleled the TGF protein expression in BAL fluid. The peak of TGF protein increase in BAL fluid between 3 and 6 weeks coincided with the initial influx of inflammatory cells in BAL fluid, but preceded histologically discernable pulmonary fibrosis that was not apparent until 8–10 weeks after irradiation. In conclusion, TGF and mRNA and protein upregulation preceded the radiation-induced pulmonary fibrosis, suggesting a pathogenetic role in the development of radiation fibrosis.     

子宫阔韧带内静脉的解剖学研究及其临床意义

子宫底和体上部的静脉汇集于子宫角处浅出,应称子宫上静脉。该静脉续为卵巢静脉。子宫上静脉1条者占30%,2条者占56.7%,3条者占13.3%。子宫上静脉与输卵管峡部中点相对处的口径是3.7±0.2mm,卵巢丛与子宫上静脉汇合后的口径为5.0±0.4mm。输卵管峡部中点与子宫上静脉的间距为6.3±0.6mm。在输卵管系膜中见有输卵管静脉汇入子宫上静脉。本文研究结果认为盆腔静脉淤血症的发生,与结扎手术中损伤子宫上静脉和输卵管静脉有关。     

心电信号预测冠心病的粗糙集方法

应用粗糙集理论对心电图波形所蕴涵的大量信息进行处理，目的是评价心电信号实测指标与冠心病的关系。通过对心电图导联I中采集的心电信号进行属性约简，降低决策表的冗余性，提取用于判别冠心病的重要属性和诊断规则。实例分析表明，粗糙集理论应用于心电信号分析，可得到清晰简明的诊断规则，有助于冠心病的临床诊断。     

干扰素γ对实验性大鼠肺纤维化、肺组织干扰素γ和转化生长因子β表达的影响

目的：研究干扰素γ（IFN-γ）对博莱霉素诱导的大鼠肺纤维化实验模型肺泡炎和肺纤维化的影响,并探讨其对肺纤维化影响的可能机制。方法：60只SD大鼠随机分为正常对照组（N组）、模型对照组（M组）和干扰素γ组（R组）,每组各20只大鼠。于造模后3、7、14、28天分批处死动物,留取肺组织,观察肺泡炎和肺纤维化的程度,测定肺组织中的羟脯氨酸含量、IFN-γmRNA、转化生长因子β（TGF-β）mRNA的表达。结果：M组、R组大鼠肺泡炎3、7、14、28天均较N组严重,7天时最重,R组14天时肺泡炎显著高于M组;M组、R组肺纤维化评分及肺羟脯氨酸含量14、28天均高于N组,于28天最重;R组肺组织IFN-γmRNA表达3、7、14、28天均显著高于M组;R组肺组织中TGF-βmRNA表达在3天显著高于M组（P〈0.05）。结论：在博莱霉素诱导大鼠肺纤维化早期给予干扰素γ并不能减轻肺部炎症和肺纤维化,可能与其促进肺组织IFN-γ、TGF-β基因表达有关。