α粒子诱发转化人支气管上皮细胞BEP2D中p53基因的突变特点

目的 观察α粒子诱发BEP2D细胞转化过程中p53基因的改变。方法 聚合酶链式反应－单链构象多态性分析（PCR－SSCP）。结果 分析发现在细胞转化过程中,p53基因发生重要改变,照射后早期传代细胞中,p53外显子7就发生碱基突变,经酶切分析为249密码子的改变,外显子5／6在照后20代细胞内开始丢失,并经Southern杂交证实,以上改变均在转化过程中持续存在;而外显子8／9未见变化。结论p53基因外显子5、6、7是α粒子对DNA损伤的重要靶位点,在α粒子诱发支气管上皮细胞转化的过程中发挥重要作用。     

女性激素相关生物指标在乳腺癌癌前病变中的表达与乳腺癌发生的关系

目的：探讨女性激素相关生物指标雌激素受体（estrogen receptor，ER）、孕激素受体（progesterone receptor，PR）、雌激素诱导基因（estrogen induced gene，pS2）在乳腺非典型增生癌变过程中的意义。方法：检测正常乳腺、乳腺单纯性增生、乳腺非典型增生及原发性乳腺癌细胞中ER、PR、pS2的表达。结果：ER、PR、pS2在26例正常对照组中阳性表达率分别为11．53％，15．38％，0；在50例乳腺单纯性增生疾病中阳性率分别为22％，28％，8％；在45例非典型增生症中阳性率分别为64．44％，73．33％，31．11％；在46例原发乳腺癌中阳性率分别为67．39％，56．52％，34．78％。结论：女性激素相关生物指标ER、PR在不典型增生细胞中检测阳性时临床上应特别重视，其发生乳腺癌的可能性增大。pS2的阳性表达可能与乳腺癌的发生也有一定的关系。     

癌基因rasP~(21)、c-myc和P~(53)在乳腺增生与乳腺癌中的表达相关性及其生物学意义

目的:探讨癌基因在乳腺增生、乳腺癌中的表达相关性和生物学意义.方法:应用S-P微波免疫组化法检测乳腺单纯性增生、不典型增生(各20例),乳腺癌(40例)中rasP~(21)、c-myc、P~(53)基因产物的表达.结果:rasP~(21)在不典型增生和癌中阳性表达率较高(60%及47.5%),C-myc和P~(53)在乳腺癌及伴有转移的癌中表达率最高(57.5%—52.5%和73.3%—73.3%),良性病变表达率极低.结论:P~(21)基因在细胞早期癌变过程中具有引发作用,可能为癌的启动基因,而c-myc、P~(53)的激活与突变和乳腺癌的发生发展及癌瘤分化,侵袭转移的生物学行为和预后密切相关.     

人脑肿瘤组织中p53基因249位密码子点突变的研究

点突变是野生型P53基因失去阻止细胞分裂和抑瘤功能的主要方式。目前在多种肿瘤中如结肠癌、肺癌、成骨肉瘤、食管癌、肝癌及乳腺癌等发现有P53基因的点突变，且175、249和273位密码子是突变热点（hot0PO小“‘。更令人感兴趣的是P53基因的突变与病人的临床分期和预后相关，具有指导临床应用的价值o－”。人脑肿瘤组织中P53基因点突变研究较少。本文用聚合酶链反应——限制性片段长度多态性（PCR——RFLP）分析法检测人脑肿瘤P53基因第7外显子249位密码子点突变情况，旨在探讨P53基因突变和人脑肿瘤的发生及转归等的关系。材料和方法1…     

乳腺癌组织PTEN基因突变的PCR-SSCP分析

目的 了解抑癌基因PTEN在乳腺癌组织中的突变频率和突变类型，探讨PTEN突变与散发性乳腺癌的关系。方法 应用聚合酶链反应一单链构象多态性分析（PCR-SSCP）方法以及DNA测序技术，扩增45例散发性乳腺癌PTEN基因的9个外显子，根据构象改变检测其基因突变情况。结果 45例乳腺癌组织中检测到1例外显子2第24位密码子由A-C的错义突变，突变使蛋白结构由甲硫氨酸突变为亮氨酸。结论 散发性乳腺癌中存在PTEN抑癌基因的突变，但突变率很低。     

胃粘膜肠化生组织YNZ22基因杂合缺失及p53基因突变、蛋白表达的研究

为探讨YNZ22基因杂合缺失（LOH）及p53基因突变、蛋白表达在胃粘膜肠上皮化生中的作用,应用PCR－RFLP、PCR－SSCP及免疫组化技术检测了不同类型肠化生组织中YNZ22基因LOH、p53基因素5－8外显子突变及其蛋白表达。结果显示,YNZ22 LOH率为19.4%（6／31）,p53突变率为29.8%(14/47),p53蛋白表达率为10.0%（6/60)。将肠化生分为Ⅰ、Ⅱ、Ⅲ型,发现Ⅲ型肠化生中p53突变率及其蛋白阳性表达率分别为57.1%(8/14)和27.8%(5/18),无显著高于Ⅰ（18.2%)、Ⅱ型肠化生（2．4％）（均P＜0.05)。p53蛋白表达与p53基因突变显著相关（P＜0.05),YNZ22 LOH与p53突变及蛋白表达无显著相关性（P＞0.05)。提示,YNZ22基因LOH及p53基因异常可能在胃粘膜肠化生的发生及其癌变中起一定作用,p53基因有可能成为胃癌早期诊断的分子指标。     

粪便中检测K-ras基因突变对老年大肠癌诊断价值的研究

探讨粪便K-ras基因检测在老年大肠癌临床诊断中的价值。收集连续就诊的23例老年大肠癌患者，20例结肠癌瘤性息肉患者及20名健康老年查体者的粪便，并从中提取DNA，应用等位基因特异性杂交技术检测粪便K-ras基因第12位密码子第1，2位碱基突变情况。结果K-ras基因突变在大肠癌患者为56．52％（13／23），明显高于正常查体者的5％（1／20）（P＜0．01），与结肠腺瘤性息肉组的30％（6／20）比较，差异无显著性意义（P＞0．05）。92．31％（12／13）的大肠癌K-ras基因突变位点发生在第12位密码子第2位碱基。研究表明，结肠癌患者组织及粪便中K-ras基因突变的检出具有良好的一致性，提示粪便中检测K-ras基因突变是一种无创性的老年大肠癌的诊断方法。     

新疆细毛羊leptin基因第二、三外显予扩增片段的多态性分析

根据leptin基因在GenBank中的已知序列设计两对引物,采用PCR—SSCP和DNA测序技术在新疆细毛羊群体中进行单核苷酸多态性（SNPs）检测分析。结果表明,新疆细毛羊leptin基因第二、三外显子都具有多态性。其中,第二外显子扩增片段上有AA和AB两种基因型,AA基因型为优势基因型;而在第三外显子有CC、CD两种基因型,以CC基因型为优势基因型。并且经过DNA测序发现,在leptin第一内含子上有两处突变．即1TrG三个连续碱基的插入和C→T的碱基替换;而在第三外显子上发生了M替换突变。     

骨髓增生异常综合征和急非淋白血病患者Ki－ras癌基因12位密码子点突变

应用ＰＣＲ技术和寡核苷酸探针杂交检测４２例患者骨髓Ｋｉ－ｒａｓ癌基因第１２位密码子点突变。结果表明，在骨髓增生异常综合征（ＭＤＳ）患者中１例ＭＤＳ（ＲＡＥＢ－Ｔ）患者（１／６）发生了ＧＧＴ→ＡＧＴ的点突变，此患者诊断后３个月转为急性白血病。在急非淋白血病（ＡＮＬＬ）患者中（３／９）检测到上述同样位点点突变，其中１例发生了ＧＧＴ→ＡＧＴ和ＧＧＴ→ＴＧＴ的双突变；７例ＡＮＬＬ缓解期患者未检测到Ｋｉ－ｒａｓ的突变；２例小细胞低血色素贫血和１例巨核细胞增生性血小板降低患者亦检测到突变。对实验结果特别是Ｋｉ－ｒａｓ癌基因的点突变在ＭＤＳ转急性白血病中的意义进行了讨论。     

肺癌P53基因突变与临床关系的研究

采用ＰＣＲ－限制性片段长度多态性分析法（ＰＣＲ－ＲＦＬＰ）检测４７例人肺癌Ｐ５３基因２４９位密码子点突变。结果显示，ＮＳＣＬＣＰ５３基因２４９密码子突变率２４．２４％（８／３３），ＳＣＬＣ突变率为０（０／１４），Ｐ５３基因２４９位密码子点突变与肺癌分期、组织分化、吸烟无关（Ｐ＞０．０５），与ＮＳＣＬＣ淋巴结转移相关（Ｐ＜０．０５）提示：Ｐ５３基因２４９位密码子突变是ＮＳＣＬＣ突变热点，是评估ＮＳＣＬＣ预后因素之一。     

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.