抗人IgK和Igλ链单克隆抗体的制备及临床初步应用

用初乳和血清IgA免疫Balb/c小鼠的脾细胞与SP_2/O和NS-1瘤细胞融合,以ELISA双抗原夹心法检测获得5株杂交瘤细胞。其中3株(1D1、5C3、11A7)分泌抗人游离和结合型λ轻链;2株(4G12、14A6)分泌抗人游离和结合型K轻链。4G12与结合型K链反应比游离K链好。这些细胞株在长达1年的培养中仍能稳定分泌特异性单克隆抗体,经液氮冻存10个月后杂交瘤细胞仍保持原有的特性。用固相抗原竞争ELISA试验表明,两个抗K链McAb是针对不同的抗原位点,而3株抗λ链则针对相同的抗原位点。用于检测副蛋白和本周氏蛋白具有很好的特异性和敏感性。把McAb4G12-HRP和1D1-HRP混合,用于检测-EB病毒早期抗原(EA)和壳抗原(VCA)抗体被证明是满意的,同时试用于抗核抗体的检测也获得了初步的结果。     

产毒大肠杆菌不耐热肠毒素单克隆抗体杂交瘤细胞系的建立及初步应用

用亲和层析纯化的产毒大肠杆菌(ETEC)不耐热肠毒素(LT)免疫BALB/C小鼠,将小鼠脾细胞与SP_(2/0)骨髓瘤细胞(1:8)在PEG(MW1000)作用下融合,采用ELISA筛选抗-LT抗体阳性孔,细胞融合率为50％,抗体阳性率为4.5％,经3～5次克隆后筛选出2株分泌抗-LT杂交瘤细胞株(2D_、2E_5),用ELISA测定杂交瘤细胞培养上清及腹水的效价,后者约为前者的100倍,高达90万～110万。通过平板免疫溶血试验和ELISA双抗夹心法分别对2株杂交瘤分泌的抗LT单克隆抗体(McAb)的特异性进行了试验,结果表     

产生抗单纯疱疹病毒单克隆抗体杂交瘤细胞系的建立

用HSV-1 SM44株免疫C×S/1小鼠的脾细胞与小鼠骨髓瘤细胞系Sp2/O融合,经HAT选择培养,细胞融合率为100％。ELISA筛选后获得53孔产生抗HSV抗体的杂交瘤阳性孔。对1A12,2A8,1G8,1D10,2C5 5株细胞系用有限稀释法做2～5次克隆化,阳性克隆率均达到100％。杂交瘤细胞系经连续传代培养4个月,仍能稳定产生单克隆抗体,冻存的细胞系经复苏后亦能稳定产生抗体。应用ELISA及IF试验证明,5种单克隆抗体均与HSV-1和HSV-2标准株呈特异性反应。ELISA测定,杂交瘤培养上清抗体效价为10~(-2)～10~(-3),制备免疫腹水效价为10~(-4)～10~(-7)。5种单克隆抗体与EBV,CMV,JEV和HFRSV无交叉反应。     

抗河流弧菌单克隆抗体的建立及初步鉴定

目的 制备高效、特异的抗河流弧菌单克隆抗体(monoclonal antibody,mAb),为海洋致病细菌的快速免疫诊断提供技术支持.方法 选用雌性BALB/c小鼠(8周龄)制备成为灭活河流弧菌免疫小鼠,利用杂交瘤细胞融合技术,制备出抗河流弧菌杂交瘤细胞株,用Giemsa染色对其进行染色体鉴定,并用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)方法筛选其与河流弧菌及其他重要海洋细菌的交叉反应特异性.结果 共获得4株抗河流弧菌mAb,均符合杂交瘤细胞染色体特点,具有良好的特异性和免疫反应性,分别命名为01H10,02F12,05F3和03G10.结论 本研究制备、筛选出抗河流弧菌的mAb,有助于建立抗河流弧菌快速免疫检测试剂盒.     

抗HBsAg单克隆抗体杂交瘤的建立与临床应用

目的 制备分泌抗HBsAg单克隆抗体杂交瘤细胞株,为建立快速诊断HBsAg的方法奠定了基础。方法 通过细胞融合建立了能稳定分泌抗HBsAg单抗的杂交瘤细胞株,利用ELISA测定单抗的特异性,并用此单抗对临床标本进行初步检测。结果 细胞株分泌的单抗可与HBsAg特异性结合,与乙肝病毒E抗原和核心抗原无交叉反应,经免疫扩散试验分析为IgG1型,将此单抗用辣根过氧化物标记后可与HBsAg阳性的病人血清反应。结论 筛选出了一株适合临床应用的单抗杂交瘤细胞株。     

过敏性紫癜肾炎患儿尿液系列蛋白检测的临床意义

目的:检测过敏性紫癜肾炎(HSPN)及过敏性紫癜(HSP)患儿尿液系列微量蛋白水平,探讨它们在两种疾病中的临床意义。方法:应用酶联免疫吸附法(ELISA)测定27例HSPN、32例HSP患儿,30例正常儿童尿液中的白蛋白(Alb)、转铁蛋白(TRF)、免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、视黄醇结合蛋白(RBP)、和N-乙酰-β-D氨基葡萄糖苷酶(NAG)、D-D二聚体8种微量蛋白含量,比较3组各微量蛋白之间有无差异,HSPN组尿液Alb与其他微量蛋白有无相关性。结果:与对照组相比,HSPN组尿液Alb、IgG、IgA、IgM、RBP、NAG、D-D二聚体排泄显著升高(P<0.01);HSP组尿液IgA、RBP排泄升高(P<0.05),Alb、TRF、IgG、IgM、NAG、D-D二聚体无显著差异(P>0.05)。与HSP组相比,HSPN组尿液Alb、IgG、IgM、D-D二聚体排泄显著升高(P<0.01),IgA、NAG亦升高(P<0.05),Alb、IgG、IgM、D-D二聚体、IgA、NAG可能为早期反映HSP肾损害的指标。HSPN组尿液Alb水平与RBP呈正相关(r=0.513,P<0.01)、NAG(r=0.436,P<0.01),与TRF、IgG、IgA、IgM、D-D二聚体无相关性(P>0.05)。结论:HSPN与HSP患儿尿液部分微量蛋白含量不同,Alb、IgG、IgM、IgA、NAG、D-D二聚体微量蛋白排泄量增加可能无创性反映HSPN的病程及向HSPN发展的临床检测指标。     

原发性高血压病患者尿微量蛋白测定与管型蛋白质分析的临床意义

用ELISA法检测微量尿白蛋白(Alb)、IgG、IgA、IgM,用酶标抗体染色作尿管型蛋白成份分析,对123例原发性高血压患者进行了观察。结果提示,高血压患者尿Al1b、IgG、IgA、IgM显著升高,与疾病严重程度、血压高低、病程长短有密切关系。治疗后可降至正常。患者管型中不含IgG、IgA、IgM与Alb。两法联合应用,有助于区别肾性与原发性高血压,并可对原发性高血压的肾功能改变进行早期预测与监护。     

地高辛特异性单克隆抗体制备的研究(摘要)

研究了应用淋巴细胞杂交瘤技术制备地高辛特异性单克隆抗体(MAb),并对抗体特性进行了分析,以探讨该抗体用于监测地高辛血药浓度及治疗严重中毒的可能性。作者用过碘酸钠法将地高辛分别偶联于牛血清白蛋白(Dig—BSA)和卵白蛋白(Dig—OVA);用Dig—OVA加Freund完全佐剂腹腔注射,免疫8周龄雌性BALB/c小鼠获取牌细胞与SP2/O骨髓细胞。以ELISA间接法筛选地高辛特异性抗体分泌阳性的杂交瘤克隆,并用有限稀释法进行克隆化。所获杂交瘤细胞系经扩大培养、液氮冻存、注入BALB/c小鼠腹腔制备MAb腹水,并测定MAb的特异性、MAb与地高辛、洋地黄毒甙、毛花强心丙、毒毛旋花子甙K和     

抗—HIV单克隆抗体的的筛选与鉴定

我们用HIV抗体阳性者血浆包被聚苯乙烯条，加入纯化的HIV组织培养抗原，通过夹心ELISA法，筛选出3株能分泌抗HIV单克隆抗体杂交瘤细胞，传代稳定。培养上清经Westernblot染色鉴定含有抗-HIVgag蛋白，夹心ELISA法测得杂交瘤细胞培养上清中单克隆抗体滴达到1：6400，接种BALB/C小鼠获腹水抗体效价为1：2430000。实验表明，用此单克隆抗体检测HIV病人血清抗体敏感、特异、稳定、重复性好，是一种良好的试剂。     

抗-HIV单克隆抗体的筛选与鉴定

我们用HIV抗体阳性者血浆包被聚苯乙烯条,加入纯化的HIV组织培养抗原,通过夹心ELISA法,筛选出3株能分泌抗HIV单克隆抗体杂交瘤细胞,传代稳定。培养上清经Western blot染色鉴定含有抗-HIV gag蛋白。夹心ELISA法测得杂交瘤细胞培养上清中单克隆抗体滴度达到1∶6400,接种BALB/C小鼠获腹水抗体效价为1∶2430000。实验表明,用此单克隆抗体检测HIV病人血清抗体敏感、特异、稳定、重复性好,是一种良好的试剂。     

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.     

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.     

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