γ—^32P标记反义核苷酸对HBV基因表达的封闭效应

人工构建特异性互补于ＨＢＶ基因ｍＲＮＡ ＳＰⅡ增强子区的１５－聚反义代脱氧核苷酸并和γ－＾３２Ｐ进行末端标记，以２μｍｏｌ／Ｌ剂量作用于ＨＢＶ基因转染的ＨｅｐＧ２细胞培养２４ｈ及４８ｈ后检测掺入２．２．１５细胞的１５－Ｓ－ａｓＯＮ分别为６９．４ｎｍｏｌ／Ｌ和７５．８ｎｍｏｌ／Ｌ；对ＨＢｓＡｇ的抑制率分别为５０％和７０％；斑点杂交检测ＨＢＶ ＤＮＡ，结果表明实验组与对照组与对照组无显著性差异。     

HBs／CD3双特异性抗体介导LAK对HBVDNA转染细胞的细胞毒作用

应用化学偶联方法合成ＨＢｓ／ＣＤ３双特异性抗体，并观察其对ＬＡＫ细胞的细胞毒性作用的影响。用化学偶联剂ＳＰＤＰ将鼠源的ＣＤ３与ＨＢｓ单克隆抗体连接得到ＨＢｓ／ＣＤ３双特异性抗体；设ＨｅｐＧ－２和Ｋ５６２细胞对照组。实验显示，ＨＢｓ／ＣＤ３双特异性单克隆抗体能显著提高ＬＡＫ细胞与２．２．１５细胞结合率；应用１２５ＩＵｄＲ释放试验检测细胞毒性，发现双特异性抗体能增强ＬＡＫ细胞对２．２．１５细胞的细胞毒性，并与抗体浓度相关，ＢｓＡｂ浓度递增，细胞毒性作用随之增加。对细胞毒性作用的特异性研究表明：加入ＨＢｓ／ＣＤ３ＢｓＡｂ后，ＬＡＫ细胞对２．２．１５细胞毒性显著高于ＨｅｐＧ－２和Ｋ５６２细胞组。结果表明，ＨＢｓ／ＣＤ３双特异性抗体具有双向识别ＬＡＫ细胞和２．２．１５细胞的特性，特异地促进ＬＡＫ细胞与靶细胞结合，发挥选择性细胞毒性作用。     

白细胞介素12真核表达载体对DNA疫苗诱导免疫应答的影响

目的 观察白细胞介素１２真核表达载体（ｐＷＲＧ３１６９）对ＨＢＶ－ＳＤＮＡ疫苗（ｐＣＲ３．１－Ｓ）诱导Ｂａｌｂ／ｃ小鼠（Ｈ－２ｄ）的特异性细胞免疫应答及其对稳定表达ＨＢｓＡｇ的小鼠肥大细胞瘤Ｐ８１５细胞（Ｐ８１５－ＨＢＶ－Ｓ）成瘤性的影响。方法 肌肉注射ＤＮＡ疫苗主白细胞介素１２真核表达载体,背部皮下接种Ｐ８１５－ＨＢＶ－Ｓ细胞,观察成瘤情况,４ｈ＾５１Ｃｒ释放法检测小鼠脾细胞ＣＴＬ活性。结果：对     

半乳糖修饰的反义硫代寡核苷酸的肝细胞导向及抗乙型肝炎病毒…

人工合成两种半乳糖多聚赖氨酸导向配体，与荧光素标记的互补于ＨＢＶｍＲＮＡ多聚腺苷酸信号部分的２１－聚反义硫代脱氧核苷酸（ＡＳＯＤＮ）电性结合，与２．２．１５细胞共同孵育，４０ｍｉｎ后配体组对ＨＢｓＡｇ和ＨＢｅＡｇ的抑制率为４１％～４７％和２８％～３１％，单纯ＡＳＯＤＮ组为１１％和５％，３ｄ后配体组为８８％～８９％和７１％～７４％，单纯组为６４％和５３％，经流式细胞分析，配体组使２．２．１５细胞荧光     

半乳糖修饰的反义硫代寡核苷酸的肝细胞导向及抗乙型肝炎病毒活性研究

人工合成两种半乳糖多聚赖氨酸导向配体，与荧光素标记的互补于ＨＢＶｍＲＮＡ多聚腺苷酸信号部分的２１－聚反义硫代脱氧核苷酸（ＡＳＯＤＮ）电性结合，与２．２．１５细胞共同孵育，４０ｍｉｎ后配体组对ＨＢｓＡｇ和ＨＢｅＡｇ的抑制率为４１％～４７％和２８％～３１％，单纯ＡＳＯＤＮ组为１１％和５％；３ｄ后配体组为８８％～８９％和７１％～７４％，单纯组为６４％和５３％。经流式细胞分析，配体组使２．２．１５细胞荧光染色率达７５．３％和８３．８％；单纯组为２４．３％。结果表明，两种人工配体均具有良好的导向ＡＳＯＤＮ入肝细胞的功能，且可以明显提高ＡＳＯＤＮ抗ＨＢＶ的活性。     

反义核酸抑制荷瘤裸鼠乙肝病毒基因表达

应用２．２．１５细胞转种裸鼠，建立荷瘤裸鼠ＨＢＶ动物模型。受种裸鼠皮下可迅速长出移植瘤并分泌ＨＢＶ多项标志物至外周血液中。应用互补于ＨＢＶｍＲＮＡ ＳＰⅡ增强子区的１５聚反义硫代寡核苷酸以２０ｕｇ／ｇ鼠体重剂量连续治疗１０ｄ，结果治疗组荷瘤裸鼠血清中ＨＢｓＡｇ、ＨＢｅＡｇ及ＨＢＶ－ＤＮＡ均受到明显抑制。证明该反义核酸在动物体内同样具有良好的抗病毒效应，为从基因水平研制新一代抗乙肝病毒药物奠定了基础     

HDV/HBV感染树鼠句肝组织中Fas/Fas配体表达的初步研究

目前已有许多实验研究证据直接表明Ｆａｓ／Ｆａｓ配体（ＦａｓＬ）介导了细胞凋亡。采用免疫组化和原位杂交技术检测ＨＤＶ／ＨＢＶ感染树鼠句肝组织中ＨＤＡｇ、Ｆａｓ／ＦａｓＬ表达;并采用免疫组化双重染色分析病毒抗原表达和Ｆａｓ／ＦａｓＬ表达之间的关系。１　材料与方法１．１　标本来源　３０份ＨＤＶ／ＨＢＶ感染树鼠句肝组织,取自本院传染科ＨＤＶ／ＨＢＶ感染的树鼠句动物,经ＥＬＩＳＡ、免疫组化和原位杂交技术证实,ＨＤＶ在肝细胞中稳定复制、表达,伴ＡＬＴ升高。１．２　检测方法　①ＨＤＡｇ、Ｆａｓ／ＦａｓＬ检测：…     

受体导向药物L－HSA－AraAMP体外抗病毒效应的实验研究

以乙肝病毒（ＨＢＶ）ＤＮＡ转染的细胞株２．２．１５细胞培养上清液中ＨＢｓＡｇ、ＨＢｅＡｇ滴度及细胞内ＨＢＶＤＮＡ中间体的变化作为药物评价指标，应用四甲基偶氮唑盐（ＭＴＴ）比色分析法测定细胞毒性。结果表明：乳糖化人血清白蛋白单磷酸阿糖腺苷（Ｌ－ＨＳＡ－ＡｒａＡＭＰ简称交联物）与ＡｒａＡＭＰ具有相似的抗病毒效应。二者对ＨＢｓＡｇ最高抑制率分别为５０％，６３％，对ＨＢｅＡｓ最高抑制率分别为６２．６％，６７．２％，５００μｇ／ｍｌ交联物与１００μｇ／ｍＩＡｒａＡＭＰ对ＨＢＶＤＮＡ有相似的抑制作用。交联物无明显细胞毒性（ＣＤ５０＞２０００μｇ／ｍｌ），ＡｒａＡＭＰ有明显细胞毒性（ＣＤ为６５８．５±１８０μｇ／ｍｌ），对ＨＢｅＡｇ抑制的选择指数分别为＞４．８，１．７。展示了受体导向抗病毒药物的良好前景。     

10到12岁玻俐维亚男孩的体格测重和肺功能

１０到１２岁玻俐维亚男孩的体格测重和肺功能［英１］／Ｖｉｌｌｅｎａ，Ｍ…／／ＩｎｔＪＳｐｏｒｔｓＭｅｄ．１９９４Ａｐｒ，１５Ｓｕｐｐｌ２．Ｓ７５－Ｂ．报告２３例ＨＡＨＳＥＳ，４４例ＨＡＬＳＥＳ，４３例ＬＡＨＳＥＳ和２８例ＬＡＬＳＥＳ男孩体格测量结果，...     

高压氧对实验兔海水型呼吸窘迫综合征的预防作用

目的　观察高压氧（ＨＢＯ）对海水淹溺肺水肿（ＰＥ－ＳＷＤ）的预防作用。方法　复制ＰＥ－ＳＷＤ动物模型后,３２ 只兔随机分为ＰＥ－ＳＷＤ对照组、药物治疗组和ＨＢＯ组。对３ 组兔的动脉血气酸碱指标、Ｃａ２＋ 沉淀反应颗粒、ｃ－ｆｏｓ ｍ ＲＮＡ和ｃ－ｊｕｎ ｍ ＲＮＡ 等进行自动检测和定量分析比较,并观察分析３组兔的存活时间和海水型呼吸窘迫综合征（ｓｅａｗ ａｔｅｒ－ｒｅｓｐｉｒａｔｏｒｙ ｄｉｓｔｒｅｓｓ ｓｙｎｄｒｏｍ ｅ,ＳＷ－ＲＤＳ）发生率。结果　ＨＢＯ组兔ＰａＯ２ ,ＳａＯ２ 和ｐＨ ３ 项指标比药物组显著升高（Ｐ＜ ０．０１）,动物存活时间［（４３．０３±７．１９）小时］比药物组［（２３．５８±１．４９）小时］明显延长,Ｃａ２＋ 沉淀反应颗粒,ｃ－ｆｏｓ ｍ ＲＮＡ,ｃ－ｊｕｎ ｍ ＲＮＡ和ＳＷ－ＲＤＳ发生率ＨＢＯ组则明显低于药物组（Ｐ＜ ０．０１）。结论　ＨＢＯ可明显提高ＰａＯ２ 和ＳａＯ２,有效改善低氧血症和代谢性酸中毒,减轻ＰＥ－ＳＷＤ时肺组织的损伤,从而可防止ＰＥ－ＳＷＤ向ＳＷ－ＲＤＳ转化。     

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.     

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）     

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.