Diagnostic use of anti-modified nucleoside monoclonal antibody.

By use of monoclonal antibodies (MoAbs) termed APU-6 and AMA-2, we determined the usefulness of urinary modified nucleosides, pseudouridine and 1-methyladenosine, as markers for malignancy. In patients with leukemia and other forms of cancer, these nucleosides elevated significantly and reflected the disease status of patients. The immunohistochemical analysis showed that cancer cells were specifically stained with the MoAbs. Chemical identification of the cellular components reactive with the MoAbs revealed that APU-6-associated antigens were mainly rRNA and AMA-2-associated antigens were mainly tRNA. These results suggest that APU-6 and AMA-2 would be useful tools for clinical and biological studies of cancer.     

Novel use of the BD FACS SPA to automate custom monoclonal antibody panel preparations for immunophenotyping

BACKGROUND: The effective and accurate diagnosis of hematologic malignancies relies on flow cytometric immunophenotyping. Selected combinations of monoclonal antibodies (mAbs) arranged in multicolor panels allow for the accurate definition of normal and abnormal hematologic cell populations. The most time-consuming and crucial step in the staining process involves dispensing combinations of multiple mAbs into their appropriate staining tubes. This step is prone to error, requires concentration and accuracy, and is dependent on technologist experience.The Becton Dickinson BioScience (BD) FACS Sample Prep Assistant (SPA) is touted as a breakthrough in automated in vitro diagnostic sample preparation. The SPA is designed to automate BD MultiTESk and BD TriTest lyse/no-wash assays. However, because most cases in our laboratory require tedious application of unique four-color mAb cocktails for leukemia and lymphoma testing, we wondered whether the SPA would be helpful in accurately dispensing these mixtures. METHODS: The mAb panels were prepared by the SPA in two separate timed runs and on separate days. Eleven specimens (nine from patients and two from normal volunteers) were split and stained with four-color cocktails created by the SPA or manually. The percentage of positive (%P) cells and mean fluorescent intensity for each mAb pair were determined. These values were plotted against each other and correlation values were calculated. To quantitate timesaving in the laboratory, two technologists prepared individually the same mAb panels and were timed. RESULTS: The correlation between the two methods was high; r(2) was 0.988 for 158 %P antigen pairs; no bias between the manual and robotic methods was detected with the Wilcoxon rank test. Bland-Altman analysis indicated no obvious relation between the difference and the mean of %P cells, suggesting that the SPA successfully dispensed antibodies for leukemia/lymphoma panels. The two methods may be interchangeable, although the limited sample size prohibits this conclusion from Bland-Altman statistics alone. In addition, one possible error was detected in the SPA-prepared panels. The SPA averaged 65 min/run, the experienced technologist 12.95 min/run, and the inexperienced technologist 54.9 min/run. CONCLUSIONS: SPA dispensing time was twice the average manual dispensing time; however, SPA use was completely automated and freed the technologist to perform other tasks. SPA use permitted preemptive preparation of mAb panels and thus streamlined processing; however, the cost of the assay and the amount of reagent waste increased. It is certain that software modifications by BD could decrease the SPA reagent dispense time and decrease the cost associated with reagent waste when the SPA is used in this novel fashion.     

单克隆抗体在肿瘤血管超声分子成像中的应用进展

超声靶向微泡的研制有力地推动了超声分子成像技术的发展.所谓超声分子成像,是指超声微泡通过理化作用修饰后可以特异性作用于病变组织的分子标志物,利用微泡的声学特性突出显示病变部位,间接反映了病变组织细胞及分子水平的异常变化[1-2].     

Detection of elevated amounts of urinary pseudouridine in cancer patients by use of a monoclonal antibody

Preparation of anti-pseudouridine monoclonal antibodies (MoAbs) and their applications for the quantitation of urinary pseudouridine in cancer patients are described. Seven MoAbs were selected. Five MoAbs were specific for pseudouridine and two MoAbs were cross-reactive with uridine. The most specific antibody, APU-6, was used in an enzyme-linked immunosorbent assay (ELISA) to determine urinary pseudouridine. Sensitivity was in the picomole range and the accuracy was nearly equal to that of the high performance liquid chromatography (HPLC) assay. The amount of pseudouridine in the urine of 28 healthy donors was 31.17 +/- 9.94 nmol/mumol creatinine. In 55% (35/63) of patients with cancer, urinary pseudouridine was elevated above the normal mean + 2 SD (51.04 nmol/mumol creatinine). Particularly, all of the patients (15/15) with leukemia and lymphoma had elevated levels of pseudouridine. These results suggest that urinary pseudouridine might be useful as a marker for leukemia and lymphoma.     

OKT3 monoclonal antibody in cardiac transplant patients

The nurse's role in caring for cardiac transplant patients involves attention to many details, including assessment of organ function, provision of care, and administration of immunosuppressive medications. OKT3 monoclonal antibody, an immunosuppressive agent under investigational use in the cardiac transplant population, requires special nursing considerations.     

Measurement of cyclosporine in plasma from patients with various transplants: HPLC radioimmunoassay with a specific monoclonal antibody compared

This study compares cyclosporin A (CsA) concentrations in plasma from patients receiving various transplants, as measured by HPLC and RIA with a monoclonal antibody for CsA and an 125I-labeled ligand. The RIA was restandardized with in-house standards because it overestimated CsA by an average of 23%. The RIA was sensitive to 2 micrograms/L, the standard curve was linear from 20 to 500 micrograms of CsA per liter, analytical recovery was 98%, and CVs were less than 8% for intra- and interassay precision. RIA (y) vs HPLC (x) for 283 plasma samples from 145 patients gave a slope = 1.1256, r = 0.979. When the results were segregated according to transplant type, CsA in liver and heart recipients was overestimated by RIA as compared with HPLC: slope = 1.202, r = 0.973 and slope = 1.1477, r = 0.983, respectively. Adult and pediatric CsA values were acceptable when RIA and HPLC were compared: slope = 1.0755, r = 0.977 and slope = 1.0563, r = 0.980, respectively. For six samples (four heart, two liver recipients) where HPLC and RIA values demonstrated wide discrepancies, repeat HPLC and analysis of eluate fractions gave CsA concentrations nearer values by the initial HPLC assay. We conclude that this RIA cannot be substituted for HPLC in the case of heart and liver recipients. The need for each laboratory to standardize the RIA is obvious.     

Therapeutic strategy for acute rejection in organ transplantation--optimal use of monoclonal antibody

Transplantation has now become the treatment of choice for end-stage organ failure. However, acute rejection still occurs in approximately 20-50% of cadaver or living donor kidney recipients during the first year post-transplantation and lead to permanent damage of graft function or loss. Recently chimeric(basiliximab) and the humanized(daclizumab) monoclonal antibodies against the alpha chain(CD25) of the interleukin 2 receptor(IL-2R) introduced in the late 1990s. Induction therapy with these antibodies results in a 15-20% reduction in acute rejection episodes after renal transplantation. The availability of anti-IL-2R monoclonal antibodies offers a further opportunity for graft-specific induction therapy.     

A monoclonal antibody against basal cells of human epidermis. Potential use in the diagnosis of cervical neoplasia.

A murine monoclonal antibody was generated against human skin cells obtained from psoriatic plaques. The antibody, called VM-2, recognizes an epitope expressed on the basal cell layer of human skin and other epithelia. VM-2 also binds to cultured cells from a variety of human carcinomas including HeLa cervical carcinoma, A-431 vulvar carcinoma, A-549 lung alveolar carcinoma, and SCL-1 skin squamous cell carcinoma cells. In several primary human cell lines, including fibroblasts, endothelial cells, and cells from the hematopoietic lineage, the antigenic site recognized by VM-2 could not be detected. The cellular antigen when immunoprecipitated by VM-2 from both normal and transformed cells appears to be proteins of approximately 100,000 and 120,000 mol weight. In frozen sections from human tumor-containing tissues, VM-2 labels skin, cervical, and lung squamous carcinoma cells, as well as skin basal carcinoma cells. Malignant cells present in exfoliative smears from epidermoid invasive neoplasias of the cervix are also selectively recognized by VM-2 in distinction to normal squamous cervical cells. VM-2 is thus directed against an antigen associated with neoplastic cells when applied in selected sites of exfoliative cytology. This monoclonal antibody represents a new reagent that should prove useful in the diagnosis of cervical neoplasia.     

用单克隆抗体定量测定糖尿病患者尿IgG_4

用特异性单克隆抗体，以免疫酶联法测定正常人及９５例糖尿病患者尿液ＩｇＧ_４，同时用放射免疫法测定尿中总ＩｇＧ，结果糖尿病患者尿液中ＩｇＧ_４水平高于正常对照组（Ｐ＜０．０１），其增高程度与白蛋白尿程度关系密切，而总ＩｇＧ于临床蛋白尿阶段才明显增高。提示尿液ＩｇＧ_４是反映肾小球损害程度较敏感的指标。     

住院肿瘤患者HCV抗体检测结果分析

目的了解肿瘤医院住院患者中丙型肝炎病毒（HCV）的感染状况。方法对本院2008年1月至2012年12月期间的住院患者用酶联免疫吸附法（ELISA）检测血清HCV抗体,筛选出HCV抗体阳性患者相关资料进行回顾性分析。结果从67438例患者中检出HcV抗体阳性327例,其中40～59岁龄段的占53．21％,男女比例为1．11：1．00,科室分布以肝胃病区为主,2011年检出率最低为4．24‰,2008年检出率最高为5．95‰,病种以肝癌最多,占23．28％。结论肿瘤患者丙肝抗体阳性率低于一般人群、年龄跨度大、科室分布广、肿瘤谱宽。因此,对患者术前、输血前和各种创伤侵袭性操作前应该对其进行HCV常规检查,以防止院内感染的发生。     

Purification of the Borrelia burgdorferi flagellum by use of a monoclonal antibody

The flagellum associated polypeptide p41 is an immunodominant antigen of Borrelia burgdorferi in the early and late stages of Lyme borreliosis. p41 was prepared by affinity chromatography using monoclonal antibody specific for p41. An immunoglobulin class specific ELISA (IgM-, IgG-ELISA) was established with purified p41 as antigen and compared to the conventional ELISA with whole cell ultrasonic antigen. Whereas the sensitivity of IgM- and IgG-ELISA was comparable in both antigen preparations, crossreactivity of sera from syphilitic patients was reduced in the p41 IgG-ELISA. Discrepant results obtained by use of ultrasonic antigen or p41 antigen, were controlled by Western blots. A correlation between the results of p41-ELISA and Western blot was shown.     

护士主导的喉癌术后患者随访清单的制订

目的 研制喉癌术后随访清单,用于在护士主导的喉癌患者术后随访过程中系统、动态地评估患者,及时发现并解决问题。方法 基于文献研究、对喉癌术后患者半结构式访谈研制随访清单初稿,采用改良德尔菲法分别对北京市、广东省、山东省、山西省的17名专家进行第1轮咨询,对第1轮中返回问卷的15名专家进行第2轮咨询,并在喉癌术后患者中测试清单。结果 2轮专家咨询的问卷有效回收率分别为88.2%和100%。第2轮咨询中,专家权威系数为0.884,一、二、三级指标专家意见的Kendall协调系数分别为0.193、0.152、0.214 （P<0.05）。最终形成的喉癌术后随访清单包括一级指标4项（生理状况、精神状况、家庭或社会状况、功能状况）,二级指标15项,三级指标42项。结论 经专家咨询确定的护士主导的喉癌术后随访清单科学、合理,能为喉癌术后患者随访过程的动态评估及制订个体化 随访方案提供依据。     

核素骨显像对食管癌术后随访的应用价值

目的 通过核素骨显像对食管癌术后患者的检查结果。探讨食管癌术后发生骨转移的高峰时间。方法 对116例食管癌术后的患者依病程分为2组，一组为术后12个月内，另一组为大于12个月，对其二组分别行核素骨显像检查，观察其结果。结果 发现食管癌术后12个月内者，核素骨显像阳性率为38.24%，高于大于12个月组，后者为14.58%；腺癌的骨转移阳性率为48.28%，而鳞癌为21.83%。结论 核素骨显像对食管癌术后1年内的监测有一定的意义。临床医生应根据患者的术后时间，病理分型，选择核素骨显像检查，以免造成漏诊。     

Quantification of lactate dehydrogenase-1 in serum with use of an M-subunit-specific monoclonal antibody

We have developed a rapid, one-step assay for measuring lactate dehydrogenase-1 (LD-1) activity in serum after extraction of LD-2, LD-3, LD-4, and LD-5 isoenzymes by an immobilized M-subunit-specific monoclonal antibody (D.8.1). In the assay, 100 microL of serum is mixed with 50 microL of a suspension of 0.8-micron-diameter latex particles coated with 30 micrograms of the monoclonal antibody D.8.1, then incubated at room temperature for 5 min. The latex particles, to which LD-2 through LD-5 are bound, are pelleted by centrifugation for 2 min at 12,000 X g, and the LD-1 activity is measured kinetically in the supernatant fluid. We optimized the assay for antibody immobilization, antibody concentration, and time and temperature of incubation. Serum bilirubin concentrations up to 0.33 g/L (0.56 mmol/L) did not interfere in the assay. Hemolysis interfered solely through LD-1 released from erythrocytes. The within-assay CV for low-concentration quality-control material (LD-1 33 U/L) was 3.5% (n = 9) and for high-concentration material (LD-1 185 U/L) was 1.9% (n = 8); the between-assay CVs for the two materials were 6.1% (n = 9) and 2.5% (n = 10), respectively. The LD-1 activity measured in 98 samples by our assay compared well with a two-step polyclonal antibody-based assay (Isomune-LD, Roche Diagnostic Systems; r = 0.998) and with an electrophoretic method (Paragon, Beckman Instruments; r = 0.956).     

Phase I study of a murine monoclonal anti-lipid A antibody in bacteremic and nonbacteremic patients.

Nine patients with suspected gram-negative bacterial sepsis were studied to determine the safety, pharmacokinetics, and immunogenicity of XMMEN-0E5, a murine immunoglobulin M monoclonal antibody directed against the core lipid A region of bacterial endotoxin. Antibody was administered by single intravenous infusion of 1 to 4 h duration at doses ranging from 0.1 to 15 mg/kg. Five patients had positive blood cultures for gram-negative bacteria, one patient had Torulopsis septicemia, one patient had gram-negative bacterial meningitis, and two patients were culture negative. No evidence of antibody-mediated toxicity was observed at any dose level. The serum half-life of the antibody was approximately 10 h at doses of 0.1 to 7.5 mg/kg and approximately 18 h at a dose of 15 mg/kg. No apparent difference in clearance of antibody was observed between bacteremic and nonbacteremic patients. Human anti-mouse antibodies were detected in the sera of three evaluable patients that received doses equal to or greater than 2.0 mg/kg but not in patients that received lower doses of antibody. This study demonstrates that XMMEN-0E5 is well tolerated at doses from 0.1 to 15 mg/kg and may be immunogenic at doses of 2.0 mg/kg and above. Controlled trials to establish the efficacy of this antibody in the treatment of gram-negative bacteremia are indicated.     

Effects of radiolabelled monoclonal antibody infusion on blood leukocytes in cancer patients

This study was undertaken to investigate the effects of a single infusion of radiolabelled murine monoclonal antibody (MAb) on peripheral blood leukocytes in cancer patients. Eleven patients with disseminated colon cancer, malignant melanoma, or lung adenocarcinoma were infused with 111In-labelled anti-ZCE 025, anti-p97 type 96.5c, or LA 20207 MAb, respectively. Blood samples were obtained before infusion, immediately after infusion (1 hr), and at 4 and 7 days postinfusion. Flow cytometry analysis of CD3+, CD4+, CD8+, CD16+, and CD19+ lymphocytes showed increasing CD4:CD8 ratios in seven patients after infusion. This phenomenon was not restricted to antibody subclass or to type of cancer. Two of the remaining patients exhibited a marked post-infusion increase in CD8+ cells. In all three patients with malignant melanoma, decreasing levels of CD16+ lymphocytes were noted after infusion and natural killer cell cytotoxicity showed fluctuations which paralleled the changes in the CD16+ subpopulation. Oxygen radical production by phagocytic cells was markedly affected in three subjects. These results suggest that a single infusion of radiolabelled murine MAb may alter the balance of critical lymphocyte subpopulations and modulate other leukocyte responses in cancer patients.     

鼻窦炎术后出院病人随访卡的应用

鼻窦炎、鼻息肉是耳鼻咽喉科常见病，多发病。随着鼻内镜外科技术的日臻成熟，鼻窦炎、鼻息肉治疗已进入新的时代。但是完成鼻内镜手术（Fess）只是鼻窦炎治疗的开始，术后定期换药至关重要。有的病人认为，手术后就能治愈，术后不愿意回院随访；术后出院病人回院复诊时由于找不到主管医生（医生要在病房、门诊轮流工作），给病人换药带来不便。针对上述问题，建立了随访卡，给每个鼻窦炎手术后病人发放随访卡，病人凭随访卡回院换药，效果良好。现介绍如下。     

1例肝脏移植术术后随访期病人的健康教育

肝移植是肝硬化、肝癌及重症肝病病人根治治疗的最佳方法。近年来在国内外医学领域迅速开展，术后5年的生存率为70％～75％。肝移植后围手术期的护理一直倍受重视，在病人安全度过围手术期以后大多数病人即可出院，进入术后随访期。由于此期病人是在家休养，就要求病人掌握正确的健康休养知识，以免出现慢性排异反应、严重的术后并发症及不正常的心理变化等，这常常被许多医院所忽视，从而影响到病人的生命质量。