错配PCR限制片段长度多态性分析：检测HBV前C区A＿（83）点突变

为建立一种简易的检测ＨＢＶ前Ｃ区Ａ８３点突变的方法，设计了一种针对Ａ８３变异株的错配引物，通过ＰＣＲ，可在Ａ８３变异株的扩增产物中引入一个ＢＳＵ３６Ｉ限制酶位点，将扩增产物（１０２ｂｐ）用该酶消化，含变异者显示８２ｂｐ的酶切片段，以此判定的变异与测序结果一致。用本法检测了慢性乙型肝炎１２例和随访ＨＢｅ血清转换者５例，结果表明前Ｃ区Ａ８３变异与病变发展一致。     

A simple and sensitive method to assess ischemia occurrence in the setting of focal cerebral ischemia in rat： A comparative study

Focalcerebralischemiamodelinducedbyintralu minalthreadembolicmiddlecerebralarteryocclusion(MCAO)inratismostwidelyusedinthestudyofis chemiccerebralvasculardisease[1] .Neurologicalevalu ationwasusuallyappliedtoselectischemicratsafteroperation[2 ] .Previousstudiesindicatedthatneurologi calevaluationwasnotalwayscorrelationtobrainis chemicinjury[3,4 ] .Mostmodelsofcerebralischemiaaresurgicallyin vasiveandthusrequireanestheticsduringpreparation .Sleepingtimeaftergeneralanesthesianotonlydependsonha…     

A novel KIAA0196（SPG8） mutation in a Chinese family with spastic paraplegia

Hereditary spastic paraplegia （HSP） is one of the most.heterogeneous genetic neurodegenerative diseases,caused by mutations in more than 50 different genes.The eighth HSP locus,SPG8,is on chromosome 8p24.13.SPG8 is a rare autosomal dominant-HSP （AD-HSP） caused by mutations in the KIAA0196 gene,with only seven SPG8families described to date.1 Here,we described the clinical characteristics of AD-HSP caused by a novel mutation in the KIAA0196 gene in a Chinese family.     

Wastewater Treatment in a Hybrid Biological Reactor （HBR） ： Nitrification Characteristics

Objective To investigate the nitrifying characteristics of both suspended- and attachedbiomass in a hybrid bioreactor. Methods The hybrid biological reactor was developed by introducing porous ceramic particles into the reactor to provide the surface for biomass attachment.Microorganisms immobilized on the ceramics were observed using scanning electron microscopy (SEM). All chemical analyses were performed in accordance with standard methods. Results The suspended- and attached-biomass had approximately the same nitrification activity. The nitrifying kinetic was independent of the initial biomass concentration, and the attached-biomass had a stronger ability to resist the nitrification inhibitor. Conclusion The attached biomass is superior to suspended-biomass for nitrifying wastewater, especially that containing toxic organic compounds. The hybrid biological reactor consisting of suspended- and attached-biomass is advantageous in such cases.     

Morphologic damage of neurons in hippocampus （CA－1） following focal brain cortex contusion： a quantitative histopathologic st

Ｍｏｒｐｈｏｌｏｇｉｃｄａｍａｇｅｏｆｎｅｕｒｏｎｓｉｎｈｉｐｐｏｃａｍｐｕｓ（ＣＡ－１）ｆｏｌｌｏｗｉｎｇｆｏｃａｌｂｒａｉｎｃｏｒｔｅｘｃｏｎｔｕｓｉｏｎ：ａｑｕａｎｔｉｔａｔｉｖｅｈｉｓｔｏｐａｔｈｏｌｏｇｉｃｓｔｕｄｙ￥ＺｈａｎｇＪｉａｎｎｉ...     

Quantitative Analysis of the Tomographic Technetium－99m MIBI （~（99m）Tc－MIBI） Myocardial Bullseye Display： Application to Diag

ＱｕａｎｔｉｔａｔｉｖｅＡｎａｌｙｓｉｓｏｆｔｈｅＴｏｍｏｇｒａｐｈｉｃＴｅｃｈｎｅｔｉｕｍ－９９ｍＭＩＢＩ（~（９９ｍ）Ｔｃ－ＭＩＢＩ）ＭｙｏｃａｒｄｉａｌＢｕｌｌｓｅｙｅＤｉｓｐｌａｙ：ＡｐｐｌｉｃａｔｉｏｎｔｏＤｉａｇｎｏｓｉｓｏｆＣｏｒｏｎａ?..     

Common non-synonymous polymorphisms in the BRCA1 Associated RING Domain （BARD1） gene are associated with breast cancer susceptibility： a case-control analysis

The BRCAI Associated RING Domain （BARDI） gene has been identified as a high penetrance gene for breast cancer, whose germline and somatic mutations were reported in both non-BRCA1/2 hereditary site-specific and sporadic breast cancer cases. BARDI plays a crucial role in tumor repression, along with its heterodimeric partner BRCAI. In the current study, we tested the hypothesis that common non-synonymous polymorphisms in BARDI are associated with breast cancer susceptibility in a case-control study of 507 patients with incident breast cancer and 539 frequency-matched cancer-free controls in Chinese women. We genotyped all three common（minor allele frequency （MAF） 〉 0.10） non-synonymous polymorphisms（Pro24Ser, Arg378Ser, and Va1507Met） in BARDI. We found that the BARDI Pro24Ser variant genotypes （24Pro/Ser and 24Ser/Ser） and Arg378Ser variant homozygote 378Ser/Ser were associated with a significantly decreased breast cancer risk, compared with their wild-type homozygotes,     

Determination of Conversion Coefficient to Evaluate Couple-year of Protection （CYP） for Tubectomy and Vasectomy in Urban and Rural Regions of Iran： IMES Study

Objective To calculate couple-year of protection （CYP） by conversion coefficient （F） of tubectomy and vasectomy in urban and rural regions of Iran. Methods A total of 103 450 married women aged 10-49 years in 2005 across urban and rural regions of lran were sampled by multi stage cluster sampling. The data were collected by household survey and direct interview and analyzed by STATA8.0 software and survey analysis commands. Results Mean age of the women at the time of tubectomy estimated 31.9 years and conversion coefficient of tubectomy was 17.1 ±0.1. Mean age of women at the time of her husband vasectomy estimated 31.48 years and its conversion coefficient was 17. 5 ± 0. 1. Cluster analysis defined different regions of Iran on the basis of evaluated conversion coefficients of tubectomy and vasectomy which both of them presented seven clusters. Literacy of women and total coverage of family planning in a region had a direct relationship with this conversion coefficient （P〈0.05）. Conclusion The variations observed in the conversion coefficients and their dissimilarity among different regions of Iran may be due to a variety of fundamental factors of which literacy and culture are of important factors.     

‘Reversing＇ the Flow of the Ancient Chinese Calendrical Subseasonal Phases Jie Qi in Australia：The Construction of a Chinese Medical ＆ Agricultural Lunisolar Calendar （Northern and Southern Hemispheres ）（ Continued ）

Jie qi（subseasonal phases or solar qi）constitutes the solar component of the ancient Chinese lunisolar calendar. It divides the year into twenty-four subseasonal phases i.e. li chun （beginning of spring）,chunfen （spring equinox）,ete. From the premodern perspective of Traditional Chinese Medicine（TCM ）,normal climate change（qi hou bian qian ）moves in harmony with the temporal flow of the q i（qi yun ）and is thus beneficial to human health.However,abnormal,abrupt weather or climatic changes like those weather features of being too cold or too hot,too windy,too humid,too dry or a combination of two or more of these weather/climatic features which occur out of season can lead to ＇seasonal diseases＇ （shi bing）or ＇diseases brought about by external influences＇（wai gan zhi b ing）or by the＇six excessive climatic q i etiological factors＇（liu yin zhi bing 六因致病）.Some examples of seasonal diseases are SARS,influenza,the common cold and heat stroke. But the temporal flow of the subseasonal qi （qi yun） in Australia is the reverse of that of the northern hemispherical regions of China,United States and Europe.This situation thus leads to a temporal desynehronisation of TCM practice and practitioners with the rhythm of the seasons.In order to address this problem,I have constructed a Chinese Medical ＆ Agricultural Lunisolar Calendar （Northern ＆ Southern Hemispheres）, which ＇reverses＇the flow of the suhseasonal phases in the southern hemisphere.Having this ealendrieal tool,Chinese medicine practitioners in Australia can ＇ reverse ＇ their clinical activities in accordance with the natural rhythm of the seasons here.     

一种新的HBV DNA rtA181T耐药突变检测方法

目的:通过巢式PCR及特异引物,建立一种新的乙肝病毒(HBV) DNA rtA181T耐药突变的PCR直接扩增检测方法?方法:巢式PCR方法第1轮扩增HBV DNA 逆转录酶rt活性域片段,第2轮PCR上游引物3′末端碱基设计为与HBV DNA rtA181T突变碱基相同的碱基,扩增出的目的基因片段,即为突变片段?利用该方法,本文检测了43例HBV DNA rtA181T变异标本,分析该方法检测的灵敏性;并比较分析低拷贝HBV DNA水平与高拷贝HBV DNA水平下,该方法与PCR-Sanger测序法检测HBV DNA rtA181T突变的一致性?结果:产物经测序证实,突变检测引物巢式PCR法能扩增出HBV DNA rtA181T耐药突变?在HBV DNA水平为24 U/?滋L?rtA181T突变株含量低至10%时,该方法仍能较好检测出rtA181T变异片段?对43例不同拷贝水平的HBV DNA rtA181T耐药变异临床标本检测显示,该方法检测灵敏性达100%,常规PCR-Sanger测序灵敏性为74%,差异有统计学意义(P < 0.05)?结论:基于巢式PCR及突变引物为基础的HBV DNA rtA181T耐药突变PCR直接检测法能较好地检测HBV DNA rtA181T耐药变异发生;对HBV DNA低拷贝水平下的rtA181T耐药变异检测,该方法灵敏性优于Sanger测序法,且有较好的特异性?这对早期发现HBV DNA耐药突变?及时更改抗HBV治疗策略具有重要临床指导意义?     

应用实时荧光PCR方法检测乙型肝炎病毒基因YMDD变异

[目的]建立一种检测乙型肝炎患者YMDD变异株的方法,并且研究应用拉米夫定治疗及未用药治疗的乙型肝炎初发患者中YMDD变异株发生的情况。[方法]应用实时荧光聚合酶链反应（PCR）方法对30例未经过任何药物治疗的乙肝初发患者,50例使用拉米夫定治疗1年患者,50例使用拉米夫定治疗两年患者和20例使用拉米夫定与干扰素联合治疗两年的患者同时进行YMDD变异株的检测。[结果]通过对熔解曲线的分析,在未经任何治疗的乙肝初发患者中检出YMDD变异株2例,占6．67％;拉米夫定治疗1年的患者中检出YMDD变异株13例,占26.00％;拉米夫定治疗两年患者中检出YMDD变异株25例,占50．00％;拉米夫定和干扰素联合治疗的患者中检出YMDD变异株9例。占45．00％。[结论]在未经任何药物治疗的乙肝患者中即存在YMDD变异株,并且随着使用拉米夫定时间的延长在不断增加,且联合用药并不能减少变异株的发生,因此对乙肝患者在用药前,及用药过程中进行YMDD变异株的检测是非常必要的。而实时荧光PCR方法具有简便、快速、灵敏、经济的特点,特别适用于临床对YMDD变异株的检测,从而及时的指导临床用药。     

e抗原阴性重症乙型肝炎患者HBV前C区热点变异研究

目的　研究ＨＢＶ信号肽区热点变异与重症乙型肝炎发生及转归的关系。方法　采用错配聚合酶链反应（ＰＣＲ）和限 制性片段长度多态（ＲＦＬＰ）分析方法检测６８例ＨＢｅＡｇ阴性的重症乙型肝炎病人（其中急性重肝６例、亚急性重肝３８ 例、慢性重肝２４例）及４４例慢性乙型肝炎患者的Ｔ１８６２、Ａ１８９６变异情况。结果　急性重肝的Ａ１８９６、Ｔ１８６２变异分别 为６６．７％（４／６）、０（０／６）;亚急性重肝４２．１％（１６／３８）、１５．８％（６／３８）;慢性重肝 ２５．０％（６／２４）、１６．７％（４／２４）;慢性肝炎 ４５．５％（２０／４４）、２．３％（１／４４）。重症乙型肝炎组与慢性肝炎组比较Ｔ１８６２变异率有显著差异（Ｐ＜０．０１）,Ａ１８９６变异率无显 著差异（Ｐ>０．０５）。结论　提示Ｔ１８６２变异与乙型肝炎的重症化密切相关。而Ａ１８９６变异与乙型肝炎重症化进程是否有 关还不能确定。     

PCR-SSCP法检测乙型肝炎病毒前C区基因变异

为探索一种快速，简便检测乙型肝炎病毒（HBV）前C区基因变异的方法，用聚合酶链反应（PCR）技术扩增94例乙肝患者血清HBV前C区基因片段，阳性扩增产物用单链构象多态性（SSCP）法进行检测，对具有不同特征性电泳图谱的PCR产物再以直接测序法加以核定，以辨别不同图谱与野生株，变异株或野生株，变异株混合感染之间的关系。结果显示：有86例患者HBV前C区PCR扩增阳性，SSCP法检测发现有3种常见的特征性电泳图谱，测序后证实，它们分别代表了野生株，变异株（nt1896位G→A）及野生株，变异株混合感染的SSCP图谱；在某些变异株图谱中，尚可见其它位点也存在变异，其中以nt1899位G→A突变最为常见，提示：以PCR-SSCP技术检测HBV前C区变异具有敏感性高，特异性强，简单，快捷，价廉等优点，而且特别适用于大样本的筛检，因此有应用价值，值得推荐。     

白细胞中乙型肝炎病毒前C区变异株的感染及动态观察

目的 :探讨白细胞中HBV前C基因变异对肝功能的损害。方法 :用特异性引物聚合酶链反应 (3’BS -PCR) ,检测 10 1例慢性乙型肝炎患者。结果 :①白细胞中变异总检出率 34.6 % ;②白细胞中存在HBV前C基因变异时血清中的谷丙转氨酶 (ALT)普遍升高。结论 :①慢性乙型肝炎患者白细胞中不仅普遍存在HBVDNA ,同时还存在HBV突变株 ;②白细胞中的变异株感染可能与发病程度有关。     

Ad-1.2乙型肝炎病毒感染HepG2细胞后HBV cccDNA的动态分析

目的 研究Ad-1.2 HBV感染HepG2细胞后HBV的复制情况.方法 携带1.2拷贝FIBV DNA的腺病毒体外感染人肝癌细胞株HepG2,ELISA法检测培养上清中HBsAg、HBeAg的动态表达;利用质粒抽提试剂盒提取细胞内cccDNA,经绿豆核酸酶处理后,用特异引物进行实时定量PCR检测.结果 Ad-1.2HBV感染HepG2细胞后,可有效表达HBsAg及HBeAg;感染后第1天即可在细胞内检出cccDNA,第4天达到高峰.结论 Ad-1.2HBV感染细胞,可作为抗病毒药物筛选和评价的模型.     

Detection of pre－C region mutants of hepatitis B virus from HBeAg negative patients in Xi＇an area of China

Ｄｅｔｅｃｔｉｏｎｏｆｐｒｅ－ＣｒｅｇｉｏｎｍｕｔａｎｔｓｏｆｈｅｐａｔｉｔｉｓＢｖｉｒｕｓｆｒｏｍＨＢｅＡｇｎｅｇａｔｉｖｅｐａｔｉｅｎｔｓｉｎＸｉ＇ａｎａｒｅａｏｆＣｈｉｎａＨａｎＪｉｅ（韩捷）；ＹａｎＹａｎ（阎岩）；ＤｉｎｇＺｈｅｎｒｕｏ（丁振...     

慢性乙型肝炎HBV前C区基因突变的临床研究

采用聚合酶链反应（PCR）及单链构象多态分析（SSCP）银染技术检测了32例HBV DNA阳性慢性乙型肝炎患者及乙肝病毒携带者的前C区基因变异。结果表明有前C区基因突变株感染者8例，突变株感染者检出率为25．0％；突变株的检出率以慢性乙肝重度最高，其次是慢性乙肝中度、轻度患者，乙肝病毒携带者最低；抗－HBe阳性患者前C区突变株检出率高于HBeAg阳性患者。提示HBV前C区突变与病情轻重及e系统血清标志有关。     

在西安地区HBeAg阴性患者中检出乙型肝炎病毒前C区基因突变株

作用ＨＢｅＡｇ阴性，抗ＨＢｅ阳性患血清提取ＨＢＶ ＤＮＡ作为模板，用前Ｃ区引物进行聚合酶链反应（ＰＣＲ），选择３份ＰＣＲ扩增阳性产物，用ＥｃｏＲＩ酶切获得长为３１２ｂｐ的基因片段。将其克至ＰＵＣ１８质粒中，转化大肠杆菌ＪＭ１０９，提取质粒，经ＰＣＲ扩增及酶切鉴定后，用双脱氧末端标记法进行基因序列测定。结果发现，在２例患ＨＢＶ ＤＮＡ克隆中存在有前Ｃ区基因突变，即１８９６位的鸟嘌呤为腺嘌呤...