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。     

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     

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...     

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。     

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，...     

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。     

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     

PERIPHERAL NERVE REPAIR IN RATS USING VASCULARIZED FROZEN IN-SITU MUSCLE AUTOGRAFT

ＰＥＲＩＰＨＥＲＡＬＮＥＲＶＥＲＥＰＡＩＲＩＮＲＡＴＳＵＳＩＮＧＶＡＳＣＵＬＡＲＩＺＥＤＦＲＯＺＥＮＩＮ┐ＳＩＴＵＭＵＳＣＬＥＡＵＴＯＧＲＡＦＴＷＡＮＧＹａｎ（王岩）１，ＺＨＵＳｈｅｎｇ－ｘｉｕ（朱盛修）１，ＨＵＮＧＬＫ２，ＬＥＵＮＧＰＣ２１．Ｄｅｐ...     

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.     

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.     

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）     

Die rechtsmedizinische Identifizierung — Bedeutung und Beweiswert —

Zusammenfassung Bei der rechtsmedizinischen Identifizierung kann die Identität im strengen Sinn allenfalls bei lebenden Personen festgestellt werden; sonst läßt sich nur von Teilen auf das Ganze (vom Untersuchungsobjekt auf die Person) schließen, wobei die verschiedenen Merkmale des Untersuchungsobjektes entsprechend der Hdufigkeit ihres Vorkommens eine unterschiedliche Beweiskraft haben. Bei der Schädelidentifizierung mit Hilfe moderner photographischer oder elektronischer Superprojektionsverfahren ergeben sich unter Berücksichtigung der Weichteildicken so viele (fiktive) Vergleichspunkte, daß bei geeignetem Vergleichsmaterial (Photographien) Identität wegen der Vielzahl übereinstimmender Bezugspunkte in den meisten Fällen evident ist.     

Imaging of abdominal nodal spread in malignant disease

This is a review of the role of imaging procedures for the assessment of abdominal and pelvic lymph nodes. The diagnosis of malignant lymphatic spread is rarely the sole purpose of imaging, because it is usually part of a general abdominal examination, most frequently with CT or US, or increasingly with MRI. These studies are often requested in order to obtain information about the situation to be encountered during surgery, or to alert the surgeon to irresectability or to unexpected metastases outside the initially planned area of exploration. In most surgically treated tumours the role of imaging for preoperative staging is limited, due either to its insufficient sensitivity or because the initial treatment is independent of the lymph node stage. Imaging is commonly used to verify treatment response to chemo- or radiotherapy and for follow-up.Correspondence to: S. Delorme     

改进的定位技术在胸部肿瘤穿刺活检中的应用

目的：探索CT-SIM三维定位系统、体模固定技术和PET-CT融合影像导引定位技术在胸部肿瘤经皮穿刺活检中的应用价值。方法：对380例胸部肿瘤患者行改进的CT定位技术下的经皮穿刺活检术。根据肿瘤的大小、深度、毗邻关系、活动度以及患者的心肺功能状态,综合运用CT-SIM系统、体模固定技术和PET-CT融合影像导引技术,为患者进行穿刺前定位。统计穿刺定位时间长度、成功率、确诊率、并发症发生情况,并与210例采用传统铅栅定位下胸部肿瘤穿刺活检的相应资料进行比较。结果：采用改进的CT定位技术的380例患者穿刺定位精确,平均定位时间（9.5 min）较传统方法（16.8 min）缩短7.3 min,活检成功率和确诊率分别是98.7%和95.3%,高于传统定位方法的93.3%和83.3%,两者差异均具有统计学意义（P〈0.05）。穿刺并发症发生率相似,气胸发生率分别为2.8%和2.9%,咯血发生率分别为11%和12.8%。结论：根据患者状态及肿瘤特点,在CT-SIM系统快速精确定位技术的基础上,综合运用体模固定技术和PET-CT融合影像导引技术,能显著缩短经皮穿刺活检的定位时间,提高活检成功率。