稳定,高效表达血小板生成素的真核细胞株的构建

目的：血小板生成素（ＴＰＯ）在治疗血小板减少症方面具有潜在的应用前景，为构建体外稳定表达ＴＰＯ的真核细胞株，进行了ＴＰＯ的稳定表达研究。方法：采用逆转录－ＰＣＲ（ＲＴ－ＰＣＲ）方法，从水囊引产的胎儿肝组织中得到ＴＰＯ ｃＤＮＡ并进行表达。结果：经过序列分析证明获得了没有错义突变的ＴＰＯ全长ｃＤＮＡ（约１ｋｂ）。在此基础上构建的ＴＰＯ的真核表达质粒在ＣＯＳ－７细胞中瞬时表达的产物能够维持并刺激ＴＰＯ     

rhTPO克隆及其在COS—7细胞中的瞬时表达

血小板生长因子（ｔｈｒｏｍｂｏｐｏｉｅｔｉｎ，ＴＰＯ）是一种正常生理状态下的造血生长因子，它刺激造血祖细胞向巨核细胞的分化及血小板的生成和释放。该实验从６月龄人胚肝脏中分离ＲＮＡ，经逆转录ＰＣＲ等步骤克隆出了ＴＰＯｃＤＮＡ，并进行了序列分析。将ＴＰＯｃＤＮＡ亚克隆至哺乳动物细胞瞬时表达载体ｐＣＭＶ４，形成了重组表达质粒ｐＣＭＶ４／ＴＰＯ。以该重组质粒转染ＣＯＳ７细胞，可测到ＴＰＯ在ＣＯＳ７细胞中的瞬时表达     

小鼠神经生长因子cDNA的克隆及核苷酸序列分析

用Ｔ４－ＤＮＡ连接酶在限制性内切酶ＳｍａＩ存在下，将神经生长因子ｃＤＮＡ片断克隆到具有双向转录启动子的载体质粒ｐＧＥＭ３Ｚｆ（＋）中。Ｓａｎｇｅｒ双脱氧链终止法测定其核苷酸顺序表明，所克隆的ＮＧＦｃＤＮＡ片断，长３６３ｂｐ，包含了成熟神经生长因子的全部编码。该重组质粒可分别用于制备ＮＧＦｃＤＮＡ探针和ＲＮＡ探针，是研究ＮＧＦ基因结构及表达调控等方面的重要工具。     

CHANGES AND CLINICAL SIGNIFICANCE OF PLASMA ENDOTHE-LIN IN TRAUMA PATIENTS WITH HEMORRHAGIC SHOCK

ＣＨＡＮＧＥＳＡＮＤＣＬＩＮＩＣＡＬＳＩＧＮＩＦＩＣＡＮＣＥＯＦＰＬＡＳＭＡＥＮＤＯＴＨＥ┐ＬＩＮＩＮＴＲＡＵＭＡＰＡＴＩＥＮＴＳＷＩＴＨＨＥＭＯＲＲＨＡＧＩＣＳＨＯＣＫＤＵＤｉｎｇ－ｙｕａｎ（都定元）１，ＧＡＯＪｉｎ－ｍｏｕ（高劲谋）１，ＷＡＮＧＸ...     

血小板生成素的研究进展

根据近年来的文献报道，概述了血小板生成素（ＴＰＯ）的研究进展。人ＴＰＯ基因长度约为６．２ｋｂ，含有６个外显子和个内含子。所有外显子－内含子的连接符合ＧＴ／ＡＧ规则。其ｃＤＮＡ克隆由１７７４个核苷酸附加一个多聚腺苷酸尾巴构成，它是一个１０５９个核苷酸的可译框架，是一种单拷贝基因。转录起始位点位于其ＤＮＡ－１９４９位的胞嘧啶外，在转录起始位占上游１．４ｋｂ的序列中没有通常哺乳动物启动子的基序和ＴＡＴＡ     

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。     

噪声对飞行员畸变产物耳声发射的影响

目的：了解噪声对飞行员畸变产物耳声发射（ＤＰＯＡＥ）的影响及其与纯音听阈的关系。方法：飞行员６６名（１２９耳）,健康青年４０名（８０耳）,采用ＧＳＩ－１６纯年务计和ＧＳＩ－６０耳声发射测试仪分别测试纯音叶阈和ＤＰＯＡＥ,刺激声强度Ｌ１＝Ｌ２＝７０ｄＢ（ＳＰＬ）,ｆ２／ｆ１＝１．２,设定取２ｆ１－ｆ２共８个的ＤＰＯＡＥ幅值。     

A Comparison in Healing of Skull Defect Repaired with Four Different Kinds of Graft Materials in Rabbits

ＡＣＯＭＰＡＲＩＳＯＮＩＮＨＥＡＬＩＮＧＯＦＳＫＵＬＬＤＥＦＥＣＴＲＥＰＡＩＲＥＤＷＩＴＨＦＯＵＲＤＩＦＦＥＲＥＮＴＫＩＮＤＳＯＦＧＲＡＦＴＭＡＴＥＲＩＡＬＳＩＮＲＡＢ┐ＢＩＴＳＤＩＮＧＺｈｅｎ－ｑｉ（丁真奇）１，ＴＡＮＦｕ－ｓｈｅｎｇ（谭富生）２，...     

人睫状神经营养因子的原核表达及其初步纯化

目的：优化条件实现人睫状神经营养因子（ｈｕｍａｎｃｉｌｉａｒｙｎｅｕｒｏｔｒｏｐｈｉｃｆａｃｔｏｒ,ｈＣＮＴＦ）ｃＤＮＡ在大肠杆菌中的高效表达。方法：ＣＮＴＦ克隆进ｐＢＶ２２０后,表达,凝胶过滤纯化,１０日龄鸡背根神经节（Ｅ１０ＤＲＧｓ）测定其生物活性。结果：ＣＮＴＦ在ＨＢ１０１中高效表达,并具有生物活性。结论：ＳＤＳ－ＰＡＧＥ观察到相对分子质量约为２４×１０３的预期表达产物,优化表达条件后发现其在ＨＢ１０１中的表达状况最好,用凝胶过滤纯化的重组蛋白能促进Ｅ１０ＤＲＧｓ的生长     

~（99m）Tc－DTPA和~（99m）Tc－HMPAO诊断脑死亡的效

~（９９ｍ）Ｔｃ－ＤＴＰＡ和~（９９ｍ）Ｔｃ－ＨＭＰＡＯ诊断脑死亡的效果比较［英］／ＳｐｉｅｔｈＭＥ…∥ＣｌｉｎＮｕｃｌＭｅｄ．－１９９４，１９（１０）．－８６７～８７２回顾分析同一天做￣（９９ｍ）Ｔｃ标记的ＤＴＰＡ和ＨＭＰＡＯ脑灌注显像诊断脑死亡的效...     

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.     

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).     

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.     

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.     

Prevalence and severity of hip and groin pain in sub‐elite male football: a cross‐sectional cohort study of 695 players

The purpose of this study was twofold: (a) to investigate the prevalence of hip and groin pain in sub‐elite male adult football in Denmark and (b) to explore the association between prevalence and duration of hip and groin pain in the previous season with the Copenhagen Hip and Groin Outcome Score (HAGOS) in the beginning of the new season. In total 695 respondents from 40 teams (Division 1–4) were included. Players completed in the beginning of the new season (July–Sept 2011) a self‐reported paper questionnaire on hip and/or groin pain during the previous season and HAGOS. In total 49% (95% CI: 45–52%) reported hip and/or groin pain during the previous season. Of these, 31% (95% CI: 26–36%) reported pain for >6 weeks. Players with the longest duration of pain during the previous season had the lowest HAGOS scores, when assessed at the beginning of the new season, P < 0.001. This study documents that half of sub‐elite male adult football players report pain in the hip and/or groin during a football season. The football players with the longest duration of pain in previous season displayed the lowest HAGOS scores in the beginning of the new season.     

Bronchial stump fistula :treatment with covered retrievable hinged metallic stents-preliminary clinical experience

Objective To evaluate the preliminaily clinical efficacy and retrievability of a retrievable hinged covered metallic stent in the treatment of the bronchial stump fistula （BSF）. Methods Between April 2003 and March 2005, 8 patients with bronchial stump fistula after pneumonectomy or lobectomy were treated with two types （A and B） of retrievable hinged covered metallic stents. Type A stent was placed in 6 patients and type B in 2 under fluoroscopic guidance. The stent was removed with a retrieval set when BSF was healed or complications occurred. Results Stent placement in the bronchial tree was technically successful in all patients, without procedure-related complications. Immediate closure of the BSF was achieved in all patients after the procedure. Stents were removed from all patients but one. Removal of the stents was difficult in two patients due to tissue hyperplasia. Patients were followed up for 6 - 21 months. Placement of the stents remained stable in all patients except one due to severe cough. Permanent closure of BSF was achieved in 7 （87.5%） of 8 patients. Conclusion Use of a retrievable hinged covered expandable metallic stent is a simple, safe, and effective procedure for closure of the BSF. Retrieval of the stent seems to be feasible. （J Intervent Radiol, 2007, 16： 253-257）