重组CFP10-PPE68融合蛋白检测结核分枝杆菌感染的初步应用

目的以结核分枝杆菌(Mycobacterium tuberculosis,MTB)RD1区特异性培养滤液蛋白10(culture filtrate pro-tein10,CFP10)与戊糖-5-磷酸-3-差异构象酶68(pentose-5-phosphate-3-epimerase68,PPE68)的融合蛋白CFP10-PPE68作为包被抗原,建立检测结核病患者血清中MTB抗体的间接ELISA法。方法采用亲和层析法纯化重组CFP10-PPE68、CFP10和PPE68蛋白;分别以3种蛋白作为包被抗原,建立检测结核病患者血清中MTB特异性抗体的间接ELISA法;采用方阵滴定法对建立的间接ELISA法的条件进行优化;并以临床诊断为金标准,对间接ELISA法的特异性、敏感性和准确性进行验证。结果纯化的重组CFP10-PPE68融合蛋白纯度约为70%。分别以重组CFP10-PPE68、CFP10和PPE68蛋白作为包被抗原的最佳包被浓度分别为2、4、4μg/ml,血清最佳稀释度均为1∶1 000,酶标二抗最佳稀释度均为1∶5 000。3种蛋白对肺外结核的诊断效果均优于肺结核,且重组CFP10-PPE68融合蛋白检测肺外结核的特异性、敏感性及准确性均最佳;3种蛋白诊断结核病的特异性较高,而敏感性相对较低。结论以重组CFP10-PPE68融合蛋白作为包被抗原建立的间接ELISA法诊断MTB感染具有较高的敏感性和特异性,特别是对难以诊断的肺外结核病的诊断有一定的参考价值。     

原位杂交试剂盒在临床诊断领域的应用

原位杂交(in situ hybridization,ISH)技术是用于细胞内检测和定位某一特定靶核苷酸的新兴分子诊断技术,随着荧光原位杂交(fluorescence in situ hybridization,FISH)和显色原位杂交(chromogenic in situ hybridization,CISH)技术的问世和发展,其在灵敏度、特异性、实验室安全性等方面均有了较大进步,并逐渐开发出商品化的试剂盒,应用于微生物检测、产前诊断、肿瘤的鉴别诊断及个体化用药等多个领域。本文将对ISH技术的概况、特点及该类试剂盒在临床诊断中的应用作一综述。     

肺癌患者GM-CSF、CEA联合检测及临床意义

目的 探讨联合检测血清、胸水中粒细胞-巨噬细胞集落刺激因子(GM-CSF)和癌胚抗原(CEA)对肺癌的诊断价值和临床意义。方法 利用放射免疫法检测32名健康人的血清及78名肺癌患者(其中腺癌28例,鳞癌27例,小细胞癌23例)血清和胸水中GM-CSF和CEA的含量。结果 肺癌各组血清GM-CSF均明显低于正常组,而各组之间血清与胸水的含量差异均无显著意义。血清中CEA含量各组均明显高于正常组,且腺癌组升高幅度最大,小细胞癌升高幅度最小,各组之间差异均有显著意义。胸水CEA含量各组变化与血清相似。血清和胸水GM-CSF的含量随临床症状的加重而逐渐降低。而CEA含量逐渐升高,相关性显著。结论 肺癌的病情严重程度不同,GM-CSF、CEA的含量也不同。检测GM-CSF、CEA对临床应用GM-CSF治疗肺癌具有指导作用。     

孕妇IgG类红细胞血型不规则抗体对早期诊断Non-ABO-HDN的意义

目的探讨孕妇IgG类红细胞血型不规则抗体对早期诊断Non-ABO-HDN的意义。方法筛查4200份孕妇血清中红细胞血型不规则抗体,阳性者检测抗体特异性、免疫球蛋白类型及效价;对检测出IgG类不规则抗体者,分娩时取脐血(或新生儿血)检测血清(浆)游离抗体,并进行红细胞直接抗球蛋白试验和红细胞放散试验,以诊断其是否发生HDN。结果4200份孕妇血清中检出红细胞血型不规则抗体44份(阳性率为1.05%),其中20份孕妇血清的抗体为IgG类或IgG及IgM混合抗体,脐血(或新生儿血)检测结果,10名婴儿发生了Non-ABO-HDN,1名发生了宫内死胎。致病的抗体特异性分布为:抗-D2例、抗-c1例、抗-E2例、抗-cE1例、抗-M4例(其中1例并有抗-A)。结论孕妇体内IgG类的Rh血型系统的抗体及抗-M易引起non-ABO-HDN,检测孕妇IgG类红细胞血型不规则抗体对早期诊断Non-ABO-HDN、鉴别引发HDN的抗体型别、评估HDN的严重程度及制定治疗方案均具有重要意义。     

产前筛查母血清标志物与产科并发症关系的研究

目的通过对孕中期(孕15～20周)母血清中游离β-hcG和AFP检测,研究血清标志物与某些产科并发症的关系,以期早期发现,早期诊断产科并发症。方法对妊娠15～20周孕妇,抽取静脉血,检测AFP和fβ-hcg。结果当游离β-hcGMoM值≥2.0时,孕中期高水平AFPMoM值与早产、低体重儿、前置胎盘、新生儿窒息有关;当AFPMoM值在0.5～2.0时,孕中期高水平fβ-hcGMoM值与羊水过少,羊水污染,过期妊娠,胎窘,胎盘植入有关;当游离β-hcGMoM值在0.5～2.0时,孕中期高水平AFPMoM值与重度子痫前期、胎膜早破、前置胎盘有关。结论孕中期高水平血清标志物与一些产科并发症如早产、低体重儿、前置胎盘、胎盘植入、胎膜早破、羊水过少、羊水污染、胎儿宫内窘迫、新生儿窒息、子痫前期有关。     

汉坦病毒核蛋白基因截短片段在大肠杆菌中表达及其产物的初步应用

目的　获得汉坦病毒重组核蛋白 ,并将其应用于血清学诊断。方法　采用PCR方法 ,从带有HTN型Z10株病毒全长核蛋白基因的质粒上扩增读码框的前 35 4bp的核蛋白基因片段。将截短的核蛋白基因片段插入表达载体pGEX 2 0T ,得到重组质粒pGEX2 0 Z10trNP ,转化大肠杆菌BL2 1(DE3) ,经IPTG诱导表达 ,表达产物经Glu tathioneSepharose 4B柱纯化 ,并进行抗原性及抗原特异性检测。结果　pGEX2 0 Z10trBP表达产物是相对分子质量约为 4 0 0 0 0的谷光甘肽转移酶 (GST)融合蛋白GST Z10trNP。经Westernblot分析 ,GST Z10trNP具有良好的抗原性。用纯化GST Z10trNP为抗原 ,用ELISA间接法检测出血热病人血清、出血热疫苗免疫的家兔及小鼠血清 ,均能很好地区分阴性和阳性血清。结论　GST Z10trNP表达量高 ,易于纯化 ,并且具有良好的抗原性及抗原特异性 ,是一种安全、廉价的诊断抗原。     

磁凝集法梅毒检测试剂检测梅毒螺旋体

目的应用磁凝集法梅毒检测试剂检测梅毒特异性抗体和反应素。方法应用磁凝集法梅毒检测试剂(Tre-ponema pallidum magnetic particle agglutination,TPMPA)检测梅毒阳性血清,分析梅毒阳性血清符合率、梅毒血清抗体滴度、假阳性检出率及交叉反应。结果用TPMPA-A、TPMPA-B试剂检测26份梅毒阳性血清样品,结果均为阳性,与省血液中心和省疾控中心的检测结果的符合率为100%;应用TPMPA-B试剂检测梅毒抗体滴度结果均较用梅毒甲苯胺红不加热血清试验诊断试剂(TRUST)检测结果高2个滴度;用TPMPA-A、TPMPA-B试剂检测正常血清,结果均为阴性,与省血液中心检测结果相符;用TPMPA-A、TPMPA-B试剂检测5份风湿病患者血清和13份慢性肝病患者血清,均未出现交叉反应。结论磁凝集法梅毒检测试剂具有良好的特异性和敏感性,操作简便、省时,可初步用于检测梅毒特异性抗体和反应素。     

重组戊型肝炎疫苗的生殖毒性试验

目的探讨重组戊型肝炎疫苗对大鼠的生殖毒性。方法将Wistar雌性大鼠分为4组:低剂量组(每只20μg HEV)、高剂量组(每只40μg HEV)、生理盐水对照组及佐剂对照组,每组30只。雌鼠于交配前给药2次,初次给药后2周进行2次给药,2次给药后1周,进行雌雄鼠合笼交配,于妊娠7和14 d进行3、4次给药。每天观察雌鼠一般临床症状;交配前每周称雌鼠体重1次,交配后每周称体重2次,每周定期测定1次摄食量;于妊娠21 d安乐死孕鼠,分离出胎仔,检测疫苗对雌鼠生殖能力、胎仔发育、胎仔骨化程度、胎仔内脏畸形的影响,取孕鼠和胎仔血液,分离血清,ELISA法检测抗HEV Ig G抗体水平。结果各组大鼠精神状态未见异常,皮毛光亮,灵敏,大小便色正成形,注射局部未见充血、肿胀、溃烂、硬结等;疫苗对孕鼠体重、摄食量均无明显影响(P0.05);各剂量组窝平均着床数、窝平均黄体数、窝平均活胎数、受孕率、活胎率、吸收胎率、死胎率、畸胎率与生理盐水对照组和佐剂对照组相比,差异均无统计学意义(P0.05);各剂量组胎仔身长、尾长、体重、胎盘重和性别比与生理盐水对照组和佐剂对照组相比,差异均无统计学意义(P0.05);各剂量组胎仔上枕骨的骨化程度均在0级和Ⅰ级正常范围以内,胸骨骨化不全率与生理盐水及佐剂对照组相比,差异无统计学意义(P0.05);各组胎仔内脏均未见畸形;孕鼠及胎仔血清中均检测到抗HEV Ig G抗体。结论重组戊型肝炎疫苗对孕鼠及胎仔安全、有效。     

β-HCG联合孕酮在异位妊娠诊断中的应用

目的评价血清β-HCG联合孕酮检测在诊断异位妊娠(Ectopic Pregnancy,EP)中的价值。方法 30例EP患者为观察组;30例宫内正常妊娠者为对照组。2组分别采用血清β-HCG联合孕酮检测。结果观察组血β-HCG及血孕酮值均低于对照组,具有显著性差异(P<0.001～0.01,表1)。结论血清β-HCG联合孕酮检测在诊断EP患者中的效果可靠,具有重要意义。     

轮状病毒Vp6蛋白的免疫荧光检测

目的检测在大肠杆菌BL21(DE3)中表达的重组轮状病毒(Rotavirus,RV)Vp6蛋白和RV SA11感染的MA104细胞中不同时间表达的Vp6蛋白及其分布规律。方法应用纯化的重组A组人轮状病毒TB-Chen株Vp6(rVp6)蛋白和RVSA11分别免疫豚鼠,制备抗血清,以抗rVp6豚鼠血清作为检测抗体,应用间接免疫荧光技术检测rVp6蛋白在大肠杆菌BL21(DE3)中的表达。分别用抗Vp6和抗RV SA11豚鼠血清检测Vp6蛋白和RV SA11株在感染的MA104细胞中的分布。结果在轮状病毒感染的MA104细胞和大肠杆菌BL21(DE3)中均能检测到轮状病毒Vp6蛋白。以抗RV SA11血清作为一抗检测感染细胞,比用抗rVp6血清检测具有更强的敏感性。结论免疫荧光试验可用于轮状病毒Vp6蛋白的检测。     

“人流血”中早孕因子的分离、纯化及鉴定

目的　分离、纯化并鉴定“人流血”中的早孕因子。方法　采用DEAE纤维素离子交换层析 ,S Sepharosefastflow离子交换层析 ,ConA SepharoseCl 4B亲和层析 ,Heparin SepharoseCl 6B亲和层析等方法进行纯化。用花结抑制实验检测出其活性。以SDS 聚丙烯酰胺凝胶电泳检测其肽图。以等电聚焦电泳法检测其等电点。结果　从人流血中分离出EPF活性样物质 ,其蛋白含量为 0 .375mg/ml,SDS PAGE见 1条均匀带 ,其相对分子质量为2 6 5 0 0 ,等电点为 6 .87。结论　本研究提取的早孕因子性质与国内外报道的从妊娠初期孕妇血清中提取的基本一致     

妊娠期肝内胆汁淤积症(intrahepatic cholestasis of pregnancy,ICP)的诊断及护理干预

目的探讨妊娠期肝内胆汁淤积症(ICP)的诊断及护理方法。方法对县级28例孕晚期妊娠期肝内胆汁淤积症的临床资料回顾性分析,诊断并进行护理干预。结果 75%以皮肤瘙痒为首发症状,黄疸及其它症状出现较皮肤瘙痒发病率低、发生晚,实验室无法进行血清肝胆酸测定,但肝功能测定、胎儿监护、胎盘功能测定均可进行,经过细心的治疗护理,23例足月产,1例早产,2例胎儿生长受限,2例胎儿宫内窘迫,无产后出血、新生儿颅内出血等,体征和生化异常在2周内恢复正常。结论早期确诊本病,积极护理干预,可降低孕产妇及胎婴儿发病率,改善妊娠结局。     

丙烯抗冲共聚产品EPF30R的开发

介绍了聚丙烯抗冲共聚产品EPF30R的工艺生产特点,并详细阐述了EPF30R重要物理性能与相关工艺控制参数的关系。通过调整TEAL/DONOR比、液相和气相聚合中的氢气浓度、气相组份和乙烯含量,使多相共聚物EPF30R具有良好的高刚性、流动性和冲击性能。     

Isolate-Specific Effect of Entomopathogenic Endophytic Fungi on Population Growth of Two-Spotted Spider Mite (Tetranychus urticae Koch) and Levels of Steroidal Glycoalkaloids in Tomato

Journal of Chemical Ecology - Entomopathogenic fungi (EPF) can be experimentally established in several plant species as endophytes. Ecological effects of EPF inoculations on plant growth and...     

Preparation and photophysical behaviors of fluorescent chitosan bearing fluorescein: Potential biomaterial as temperature/pH probes

A novel method for attaching fluorescein (via its epoxy derivative) to water‐soluble chitosan, and the temperature/pH‐sensitive qualities of fluorescence were investigated. 3‐Epoxypropoxy fluorescein (EPF) was firstly synthesized by the reaction between fluorescein and epichlorohydrin, and then water‐soluble chitosan bearing fluorescein (CS‐EPF) was prepared via ring‐opening reaction with EPF. They were characterized by the methods of ¹H NMR, MS, IR, UV–Vis, and luminescence spectra, respectively. The chemiluminescent and photophysical behaviors of EPF and CS‐EPF were studied in detail. The results showed that the fluorescent chitosan could still provide temperature and pH sensitivities similar to that of fluo‐ rescein, achieving better long‐term stability and fast equilibrium response. CS‐EPF had an excellent linear response between relative fluorescence intensity and temperature in the range of 0–60°C and two linear relationships between relative fluorescence intensity and pH in 0.0–4.14 and 8.15–13.20, respectively. This investigation may provide a convenient way to prepare low‐cost and multifunctional macromolecule biomaterial to probe pH and temperature in biological systems. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 3960–3966, 2007     

共聚聚丙烯EPF30R作汽车保险杠专用料研究

在共聚聚丙烯新产品EPF30R中加入少量的EPS30R为基体对脂，用POE（辛烯-乙烯共聚物）为增韧剂、PP-g-MAH为相容剂、滑石粉为填料制得性能符合要求的汽车保险杠专用料，并研究了增韧剂、相容剂、无机填料的含量对共混材料性能的影响。结果表明：EPF30R/EPS30R质量比为85/15，POE用量为15份，相容剂用量10份，无机填料为12份时，专用料的性能达到国内外同类产品的指标。     

Novel Biomolecules in the Pathogenesis of Gestational Diabetes Mellitus

Gestational diabetes mellitus (GDM) is one of the most common metabolic diseases in pregnant women. Its early diagnosis seems to have a significant impact on the developing fetus, the course of delivery, and the neonatal period. It may also affect the later stages of child development and subsequent complications in the mother. Therefore, the crux of the matter is to find a biopredictor capable of singling out women at risk of developing GDM as early as the very start of pregnancy. Apart from the well-known molecules with a proven and clear-cut role in the pathogenesis of GDM, e.g., adiponectin and leptin, a potential role of newer biomolecules is also emphasized. Less popular and less known factors with different mechanisms of action include: galectins, growth differentiation factor-15, chemerin, omentin-1, osteocalcin, resistin, visfatin, vaspin, irisin, apelin, fatty acid-binding protein 4 (FABP4), fibroblast growth factor 21, and lipocalin-2. The aim of this review is to present the potential and significance of these 13 less known biomolecules in the pathogenesis of GDM. It seems that high levels of FABP4, low levels of irisin, and high levels of under-carboxylated osteocalcin in the serum of pregnant women can be used as predictive markers in the diagnosis of GDM. Hopefully, future clinical trials will be able to determine which biomolecules have the most potential to predict GDM.     

Role of the Macrophage Migration Inhibitory Factor in the Pathophysiology of Pre-Eclampsia

Proinflammatory cytokines are produced in pregnancy in response to the invading pathogens and/or nonmicrobial causes such as damage-associated molecules and embryonic semi-allogenic antigens. While inflammation is essential for a successful pregnancy, an excessive inflammatory response is implicated in several pathologies including pre-eclampsia (PE). This review focuses on the proinflammatory cytokine macrophage migration inhibitory factor (MIF), a critical regulator of the innate immune response and a major player of processes allowing normal placental development. PE is a severe pregnancy-related syndrome characterized by exaggerated inflammatory response and generalized endothelial damage. In some cases, usually of early onset, it originates from a maldevelopment of the placenta, and is associated with intrauterine growth restriction (IUGR) (placental PE). In other cases, usually of late onset, pre-pregnancy maternal diseases represent risk factors for the development of the disease (maternal PE). Available data suggest that low MIF production in early pregnancy could contribute to the abnormal placentation. The resulting placental hypoxia in later pregnancy could produce high release of MIF in maternal serum typical of placental PE. More studies are needed to understand the role of MIF, if any, in maternal PE.     

共聚聚丙烯EPF30R的结构性能表征及应用研究

准气相共聚聚丙烯产品EPF30R具有良好的力学性能和加工流动性，以其为基础树脂通过改性可制得多种性能符合要求的注塑产品。