应用FQ-PCR检测孕妇外周血中胎儿细胞含量

合成一对染色体单拷贝基因GAPDH特异的引物和一对Y染色体单拷贝基因SRY特异的引物,并合成两条特异性TaqMan探针,加等量的DNA到含有上述两个引物探针的PCR反应体系中,行荧光定量PCR,获得两种基因的定量值,通过这两种基因含量的比值,计算孕妇外周血中胎儿细胞含量.在30 μL FQ-PCR反应体系中,加入最少约0.05 ng 正常男性基因组DNA能够稳定地获得扩增曲线;在60例临床孕妇外周血标本DNA 的DCFQPCR扩增中有35例出现SRY扩增阳性,阳性率为58.3%;在这些标本中,GAPDH 基因Ct值范围在20~22之间;SRY 基因Ct值范围在33~40;经计算,胎儿细胞含量范围在1/10.4~1/10.6.     

动物性饵料对养殖水体中菌数和理化因子的影响

向大海马(Hippocampus kudaB leeker)养殖池中投喂动物性饵料桡足类、杂碎鱼虾等饵料后,亲海马实验组、幼海马实验组以及空白对照组水体环境因子参数差异明显,投喂杂碎鱼虾的亲海马组与投喂桡足类的幼海马组的细菌数量均高于空白对照组。3组处理后表层异养菌平均数量变动范围分别为4×103~1.78×105CFU/mL,2.5×103~7.2×104CFU/mL,2×102~5.5×103CFU/mL,而底层异养菌则分别为6.53×103~1.59×105CFU/mL,1.5×103~7.5×104CFU/mL,5×102~1.0×104CFU/mL。表层弧菌平均数量变动范围分别为55~4.08×103CFU/mL,85~7.19×103CFU/mL,10~123 CFU/mL,而底层弧菌则分别为1.13×102~7.48×103CFU/mL,2.66×102~1.16×104CFU/mL,10~175 CFU/mL。差异显著性分析结果表明,亲海马组与幼海马组细菌数量差异均不显著(P>0.05)。而亲海马组与空白组除表层弧菌数量差异不显著外,其余,底层弧菌、表底层异养细菌数量呈显著性差异(P<0.05)。空白组与幼海马组细菌数量均呈显著性差异(P<0.05)。3组池水温变化相差不大,亲海马组、幼海马组中NH4 -N含量呈升高之势,DO极不稳定,有机残留物含量、细菌数量增加趋势,导致水质恶化。     

水城县结核病流行病学调查

2000年6月,按全国结核病流行病学调查实施方案,对水城县比德乡中心村1329人进行结核病流行病学调查,共采集痰标本116人份,其中痰标本镜检阳性11人份,阳性率为9.84%,2000年12月按照市卫生局及市防疫站的安排,对比德乡瓦厂村1042人进行结核病流行病学调查,采痰10人份共计30份痰标本（每人份采集即时痰,过夜痰和次日晨痰各1份,合计每人份3份痰标本）送市防疫站进行镜检和痰培养检验,结果无1份阳性。在对此德乡瓦厂村调查的同时另外一组人员对水城县红岩乡窑上村1153个进行结核病流行病学调查,采集痰标本42人份共计126份痰标本,送市防疫站进行镜检和痰培养检验,结果镜检阳性1人,阳性率为2.38%,痰培养阳性6人,阳性率为14.28%。     

HCV不同基因型对慢性丙型肝炎抗病毒治疗疗效的影响

目的探讨干扰素联合利巴韦林治疗慢性丙型肝炎患者临床疗效的相关影响因素.方法对110例慢性丙型肝炎患者进行PEG-IFNα-2a 180μg/周联合RBV 800~1200 mg/d的标准治疗,并完成治疗后24周随访,同时分析HCV基因型、HCV-RNA定量对持续病毒学应答(SVR)的影响.应用罗氏试剂检测丙肝病毒定量,应用反转录-套式聚合酶链反应(PT-PCR)法检测HCV基因型.结果在110例进行标准联合治疗、并完成治疗后24周随访的慢性丙型肝炎患者中,68例患者获得SVR.其中,HCV基因1b型感染者SVR率为51.4%(37/72),HCV基因N-1b型感染者SVR率为81.5%(31/38);基线HCV-RNA水平1.0×10~5IU/m L的患者SVR率为71.2%(42/59),基线HCV-RNA水平≥1.0×10~5IU/m L的患者SVR率为50.9%(26/51).结论非1b基因型SVR率高于1b基因型;基线低病毒RNA水平(血清HCV-RNA水平1.0×10~5IU/m L)SVR率高于基线高病毒RNA水平(血清HCV-RNA水平≥1.0×10~5IU/m L).     

小鼠微小病毒( MVM) 荧光定量PCR 检测方法的建立及初步应用

摘要: 目的建立小鼠微小病毒( MVM) 荧光定量PCR 方法,并初步应用于实验小鼠中。方法根据NCBI 上发表的MVM( NC_001510) 基因组序列,设计引物和探针,建立MVM 荧光定量PCR 方法。考察引物探针的最佳线形范围,特异性及灵敏度,并使用该方法对178 份清洁级小鼠粪便DNA 样本进行检测。结果MVM 荧光定量PCR 方法最佳线性范围为109 ～ 104 拷贝/μL,标准曲线的线性关系良好,R2 值可达0. 99,灵敏度为101 拷贝/μL,特异性强,与大鼠细小病毒H-1 株和KRV 株、猪细小病毒均无交叉反应。应用荧光定量PCR 方法对178 份小鼠粪便DNA进行检测,结果均为阴性。结论建立的MVM 荧光定量PCR 方法具有特异、灵敏的特点,为MVM 的流行病学调查、检测提供了有效的技术支持。     

河北省滦县牛梨形虫的分子检测

本研究旨在检测河北省滦县牛梨形虫的感染情况。采集滦县牛血液样本28份,并提取基因组DNA;根据已发表的梨形虫Tams1,P32表面蛋白以及18S rRNA基因序列,设计6对梨形虫特异性引物,以牛血液基因组DNA为模板,对不同梨形虫进行PCR或巢式PCR扩增。结果表明,在28份DNA样本中检测出双芽巴贝斯虫阳性样本7份,阳性率为25.00%;其他牛巴贝斯虫阳性样本3份,阳性率10.71%;而环形泰勒虫和瑟氏泰勒虫均未检测到阳性样本。     

江苏地区无偿献血人群HCV流行病学调查

本文旨在研究江苏地区无偿献血人群的HCV感染情况及其流行特征.对2013年-2014年度江苏省血液中心157,119名健康无偿献血者血液标本进行抗-HCV ELISA双试剂检测和HIV、HBV和HCV6-混样核酸检测,对HCV RNA阳性标本用定量PCR方法检测病毒滴度情况;对HCV RNA阳性标本进行巢式PCR扩增核心区片段,根据测序结果分析基因型,分析HCV基因型的分布及其与性别、年龄的关系.结果共检出347例抗-HCV反应性标本,阳性率为0.22%.经核酸检测,在抗-HCV阴性和单试剂阳性标本中均未检出HCV RNA阳性标本;125例抗-HCV双试剂阳性标本中HCV RNA阳性标本46例,病毒血症阳性率为36.8%.巢式PCR扩增HCV RNA阳性标本并成功测序和分型36例.基因型分型为1a(2.6%)、1b(74.4%)、2a(10.2%)、3b(5.1%)、6a(5.1%)和6n(2.8%)4种基因型和6种基因亚型,未发现4型和5型.1型和2型在男性组中比例明显高于女性组,但差异无统计学意义;1型与2型患者平均年龄小于3型和6型患者,差异有统计学意义.结果显示,江苏地区无偿献血人群抗-HCV阳性率呈现下降趋势,HCV的感染与性别和职业受教育程度没有相关性,但与年龄和职业具有相关性,感染的HCV基因型以1b为主,不同基因型在不同年龄之间具有显著差异.     

沙滤对白斑综合症病毒和细菌的过滤效果

研究了沙滤对游离白斑综合征病毒(white spot syndrome virus,WSSV)和细菌的过滤效果.以凡纳对虾、Litopenaeus vannamei作为感染对象,用游离病毒2.4×105拷贝/mL浸泡感染对虾(感染初期投喂人工配合饲料),17 d内死亡率100%,第1尾对虾死亡时间为36.5 h,最后1尾死亡时间为402 h,平均死亡时间124 h;相同浓度的游离病毒经沙滤后人工浸泡感染对虾,16 d内死亡率100%,第1尾受感染对虾60h死亡,最后1尾死亡时间为374 h,平均死亡时间168.6 h;病毒沙滤后平均死亡时间延长26.8%.1.2×104拷贝/mL游离病毒浸泡感染对虾,18 d内受感染对虾死亡率100%,第1尾死亡时间为111 h,最后1尾死亡时间为415 h,平均死亡时间274.7h;相同浓度游离病毒经沙滤后浸泡感染组养殖18 d,仅1尾死亡,时间为448 h.1.2×103拷贝/mL病毒组和1.2×102拷贝/mL病毒组直接感染和沙滤后感染养殖20 d,均未出现死亡.对细菌沙滤效果研究结果表明,从2003年8月11日-12月28日连续27次检测,细菌过滤率最低为30%,最高为98.14%,平均为63.33%;其中弧菌最低为5.26%,最高为97.2%,平均为66.46%.研究结果认为沙滤对游离病毒和细菌过滤有一定的作用.为防止对虾从海区海水污染感染病毒病和细菌病,建议对虾养殖过程中,外海水采用沙滤加蓄水池中间沉淀消毒以保证养殖用水的安全.     

秋冬季凡纳滨对虾养殖池塘细菌的数量动态

2007年9月～2008年2月,研究了珠海市斗门区凡纳滨对虾4个低盐度池塘水体、沉积物中异养细菌、弧菌、芽孢杆菌数量动态,结果表明,水体中异养细菌、弧菌、芽孢杆菌在养殖前期不稳定,中后期趋于稳定,数量波动范围分别为1.8×103～1.2×105/mL、1.4×102～4.0×103/mL、1.6×102～1.7×103/mL,水体中异养细菌数量平均为1.95×104/mL;沉积物中异养细菌较稳定,弧菌呈先升后降趋势,芽孢杆菌呈上升趋势,三者的数量均高出水体2～3个数量级,数量波动范围分别为1.0×106～2.2×107/g、7.0×103～4.7×105/g、4.7×106～2.4×106/g,异养细菌数量平均为9.16×106/g     

荧光定量PCR检测HBV DNA及临床意义

目的:了解HBV感染者的常见血清学组合的HBV DNA阳性情况,探讨其临床意义。方法:对260份血清同时用荧光定量PCR与ELISA方法进行检测并进行评价。结果:40例HBsAg(+)/HBeAg(+)/抗-HBc(+)组血清HBV DNA全部阳性,阳性率为100％,含量为10~(7.62±0.69)cps/mL;110例HBsAg(+)/抗-HBe(+)/抗-HBc(+)组血清58例阳性,阳性率为52.7％,含量为10~(4.64±1.31)cps/mL;35例HBsAg(+)/抗-HBe(-)/抗-HBc(+)组血清20例阳性,阳性率为57.1％,含量为10~(4.93±1.42)cps/mL;75例HBsAg(-)组血清中,5例HBV DNA阳性,阳性率为6.7％,含量为10~(3.68±0.46)cps/mL。结论:乙型肝炎患者HBV荧光定量检测是病毒复制最直接可信的指标,动态荧光定量PCR检测HBV DNA,对动态监测乙型肝炎感染的病原学变化及判断有关药物抗-HBV复制是否有效具有一定的临床意义。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

Jane Eyre——A Feminist In The 19th Century

In the 19th century the society was controlled by men, and women were just appendants of them, they had not any rights and freedom. But Jane was an exception, she showed some characteristics of early feminist. Jane showed her characteristics of feminism in three aspects: rebellion, equality, and independence. These characteristics were helpful to her success, and feminism is the only way out for women of that time.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-