瘢痕疙瘩Fas基因(外显子6～9)突变的检测

瘢痕疙瘩是损伤愈合过程的结果 ,它们以超过原损伤范围的大量瘢痕组织形成为特征 ,而导致这种过度增殖的原因至今未明。近年来其它领域的研究证明 :Ｆａｓ系统对控制细胞增生与凋亡起重要作用 ,Ｆａｓ基因的体细胞突变与白血病、多发性骨髓瘤等疾病的发生、发展有着密切的关系。瘢痕疙瘩属良性肿瘤 ,是否也存在Ｆａｓ基因突变国内外尚未见报道。我们先前的实验研究提示了这种突变存在的可能性。本研究目的是检测瘢痕疙瘩成纤维细胞Ｆａｓ基因外显子 69是否存在突变及其突变位点。方法 :取 1 5例瘢痕疙瘩成纤维细胞进行ＰＣＲ ＳＳＣＰ检测 ,…     

p53基因cDNA突变及其蛋白高表达与大肠癌预后关系

１００份大肠癌组织，用ＲＴ－ＰＣＲ－ＳＳＣＰ检测Ｐ５３基因ＣＤＮＡ突变；ＰＡｂ１８０１单抗免疫组化检测Ｐ５３基因蛋白搞表达并对大肠癌术对后病人作５年生存随访，比较上述２结果与大肠癌预后的关系，１００例大肠癌中，ＲＴ－ＰＣＲ－ＳＳＣＰ显示５１例大肠癌Ｐ５３基因ＣＤＮＡ突变，ＰＡｂ１８０１阳性率６２％，Ｐ５３基因ＣＤＮＡ突变和Ｐ５３基因蛋白高表达与Ｄｕｋｅｓ分期无关，Ｐ５３基因ＣＤＮＡ突变与Ｐ５３基因     

小鼠内皮抑素基因克隆、表达及其抑瘤活性鉴定

利用ＲＴ－ＰＣＲ法从小鼠肝细胞扩增出Ｅｎｄｏｓｔａｔｉｎ基因,重组人ｐＵＣ１９质粒。经测序后,将片段重且人大肠杆菌表达载体ｐＤＨ并用温度诱导表达。将表达的蛋白经纯化及复性后直接注入大鼠Ｃ６胶质瘤模型内,检测其对胶质瘤生长的抑制作用。结果,在小鼠肝组织中成功扩增到Ｅｎｄｏｗｓｔａｔｉｎ基因,测序与报道列一致重组进一致。重组进ＰＤＨ后经温度诱导表达,在ＳＤＳ－ＰＡＧＥ电泳上得到一条新的蛋白表达带,分子     

急性运动对骨骼肌肌浆网钙转运功能的影响

肌浆网（ＳＲ）在调节骨骼肌收缩一舒张过程中发挥重要作用。我们在本研究中采用持续运动方式观察急性运动对ＳＲ钙转运能力的影响，发现运动后股四头肌ＳＲＣａ２＋－Ｍｇｚ＋－ＡＴＰａｓｅ水解活性下降１８．３１％（Ｐ＜０．０１），ＳＲＣａ２＋－ＡＴＰａｓｅ水解活性下降１８．４７％（Ｐ＜０．０１）；ＳＲＣａ２＋转运能力明显下降，表现为ＳＲＣａ２＋初始摄取率和ＳＲＣａ２＋最大转运能力分别下降５７．７９％（Ｐ＜０．０１）和５５．９８％（Ｐ＜０．０１）。本研究结果提示急性运动后ＳＲＣａ２＋摄取功能下降可能与运动性骨骼肌疲劳密切相关。     

用PCR及Southern杂交方法对HBsAg阴性献血员HBV感染的检测

用ＰＣＲ与Ｓｏｕｔｈｅｒｎ杂交（ＳＢＨ）方法对７８例ＨＢｓＡｇ阴性献血员进行ＨＢＶ感染检测，结果２份标本经ＰＣＲ后直接在２５４ｎｍ紫外灯下观察就可见到特异扩增带，另有１２份标本经Ｓｏｕｔｈｅｒｎ转膜杂交后可观察到特异扩增带，总阳性率较原来ＨＢｓＡｇ检测提高１７．９％，说明ＰＣＲ－ＳＢＨ可以区分ＨＢｓＡｇ阴性但仍然存在ＨＢＶ潜伏感染的献血员，使用ＰＣＲ－ＳＢＨ方法检测的ＨＢＶ可以提高输血的安全性。     

孕妇血清和胚胎组织中微小病毒B19 DNA的检出

作者建立了检测人微小病毒Ｂ１９（ＨＰＶ－Ｂ１９）的ＰＣＲ方法。两对引物（Ｐ１Ｐ２和Ｐ３Ｐ４）的设计均位于编码ＨＰＶ－Ｂ１９衣壳蛋白ＶＰ１基因区内，扩增产物分别为７００ ｂｐ和１０４ ｂｐ。比较Ｐ１Ｐ２－ＰＣＲ和Ｐ３Ｐ４－ＰＣＲ，二者均具有高度特异性和第三性，ＨＰＶ－Ｂ１９ ＤＮＡ最小检同量分别为５ ｆｇ。应用Ｐ１Ｐ２－ＰＣＲ对３２份孕妇血清和２１份胚胎组织标本进行了ＨＰＶ－Ｂ１９ ＤＮＡ的检测，结     

PCR技术在鉴定阳性重组子中的应用

介绍了采用ＰＣＲ技术在人粒细胞集落刺激因子（ｈＧ－ＣＳＦ）、人促红细胞生成素及中国人γ－干扰素（ｈＩＦＮ－γ）ｃＤＮＡ重组质粒的构建过程中快速鉴定阳性重组子的方法。经ＰＣＲ扩增鉴定为阳性的重组子，提取质粒ＤＮＡ经重组位点相应的ＤＮＡ内切酶双酶切鉴定表明，全部含有插入片段。     

多药抗药基因mdrl探针的基因克隆

多药抗药基因（ｍｕｌｔｉｄｒｕｇｒｅｓｉｓｔａｎｔｇｅｎｅ，ｍｄｒｌ）的表达水平与细胞的耐药性直接相关，检测ＭＤＲ１的表达水平可预测化疗的效果以及预后。用分子原位杂交的方法可检测单个细胞中ＭＤＲ１的表达水平。采用ＰＣＫ扩增方法获得了一段特异的ＤＮＡ片段，并将其克隆到ｐＵＣ１８载体中，经ＤＮＡ序列分析证明与文献一致。此探针可用于临床标本的分子杂交检测。     

登革2型病毒NS3基因的扩增及序列测定

采用热酚法从登革２ 型病毒４３ 株（Ｄ２ ４３） 感染的Ｃ６／３６ 细胞中提取了病毒ＲＮＡ,以病毒ＲＮＡ 为模板,进行Ｄ２ ４３ 株ＮＳ３ 基因ｃＤＮＡ 片段的反转录 ＰＣＲ 扩增,片段长度为１１７６ ｂｐ。将扩增的ｃＤＮＡ 片段克隆到Ｔ 载体ｐＢｌｕｅｓｃｒｉｐｔ ｋｓⅡ（ ＋ ） 中。通过双脱氧法测定了ｃＤＮＡ片段序列,与国际标准株ＮＧＣ株序列一致。     

α 粒子诱发人胎气管永生化纤维细胞恶性转化相关基因的分析

目的探索辐射诱发细胞恶性转化相关基因。方法本实验利用３’端引物ＨＴ１１Ｍ（Ｇ，Ｃ，Ａ）与５’端引物ＨＡＰ１７～２４所配的２４对引物同时对ＳＨＴＦ（ＳＶ４０ｉｍｍｏｒｔａｌｉｚｅｄｈｕｍａｎｆｅｔａｌｔｒａｃｈｅａｌｆｉｂｒｏｂｌａｓｔＳＨＴＦ）细胞和经２３８Ｐｕα粒子照射后能在软琼脂上形成克隆的此细胞（αＳＨＴＦ）的ｍＲＮＡ进行反转录ＰＣＲ扩增，６％测序胶电泳，放射自显影。结果发现有２３条基因片段在αＳＨＴＦ细胞中表达，ＳＨＴＦ细胞中不表达。回收差异的基因片段，随机取其中１２条重新扩增后得到６条扩增片段。其中Ｃ１７－５，Ｃ２３－１两片段经Ｎｏｒｔｈｅｒｎｄｏｔ杂交后，均与αＳＨＴＦ细胞ＲＮＡ有强阳性杂交信号，与ＳＨＴＦ细胞ＲＮＡ无杂交信号。结论本实验证实了ＳＨＴＦ和αＳＨＴＦ细胞的基因表达有差异存在。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.