Comparison of inhibitory and binding characteristics of an antibody causing acquired von Willebrand syndrome: an assay for von Willebrand factor binding by antibody

An acquired inhibitor of von Willebrand factor (vWF) activity occurring in a patient with benign gammopathy and von Willebrand syndrome (vWS) has been partially characterized. The inhibitor-induced syndrome resulted in low to undetectable plasma levels of vWF/ristocetin, vWF/botrocetin, FVIIIR:Ag, and FVIII:C with a normal to slightly prolonged bleeding time. Platelet vWF was normal. Intensive and continuous infusion of a heat-treated factor VIII concentrate (Hemofil- T, Hyland, Glendale, Calif) elevated the FVIII:C plasma levels to about 100%, with an increase in FVIIIR:Ag levels to about 340% and vWF/ristocetin levels to about 40%, much lower than expected based on the dose of Hemofil-T and its content of vWF and FVIII:C activities. The inhibitor bound to staphylococcal protein A (SpA) with high affinity, indicating an IgG antibody (Ab). An assay for the vWF-binding capacity was developed on the basis of absorption of the Ab from serially diluted plasma by SpA and removal of vWF and FVIII:C activities from normal plasma by the SpA-Ab complex. The Ab-binding site was on the vWF component of the factor VIII complex. The Ab was unable to bind isolated FVIII:C. The combined use of the new vWF- binding assay and a battery of tests for inhibition of vWF-dependent platelet aggregation with ristocetin (which detects high molecular weight vWF), with botrocetin (which detects high and low molecular weight vWF), and with platelet-aggregating factor (which detects high molecular weight vWF) provided a means of analysis of Ab effect on in vitro vWF function. Using these tests, a comparison was made of the effects of the vWS Ab with those of an Ab inhibitor occurring in homozygous von Willebrand's disease. The Ab of the vWS patient had weak inhibitory action on vWF/ristocetin without having an effect on vWF/botrocetin and platelet-aggregating factor, a high titer vWF- binding capacity, and no anamnestic response following concentrate therapy. These findings contrasted with those of the Ab occurring in inhibitor von Willebrand's disease in which vWF inhibitor and binding values were similar, with a strong anamnestic response. The findings indicate that the vWS Ab binds to an epitope on the molecular vWF in such a way that causes only limited inhibition of vWF/ristocetin function and no inhibition of vWF/botrocetin function, suggesting that these two functional domains are at separate sites.     

Subtypes of Epstein-Barr virus (EBV) in Hodgkin's disease: association between B-type EBV and immunocompromise [see comments]

Epstein-Barr virus (EBV) has been associated with Hodgkin's disease (HD) in up to 50% of cases, but the subtype of EBV involved has only recently been studied. In this report, biopsy samples from 30 patients with HD were assessed for EBV sequences using both the polymerase chain reaction (PCR) and in situ hybridization (ISH). EBV sequences were localized to the malignant Reed-Sternberg cells and their mononuclear variants (Hodgkin's cells) in 9 of the 30 cases, with 7 demonstrating A- type and 2 B-type EBV sequences. Both of the patients with B-type EBV- associated HD had features to suggest pre-existing immune compromise: one was infected with human immunodeficiency virus (HIV) and had severe CD4+ T-lymphocyte depletion; the other was a debilitated elderly patient with dementia. A previous study suggested that A-type EBV alone is associated with HD and the finding of predominantly A-type EBV in the present series is in keeping with this report. The presence of B- type EBV in the HD of patients with pre-existing immunodeficiency, taken together with the recent report that B-type EBV occurs in HIV- associated non-Hodgkin's lymphoma, suggests that B-type EBV may be an important human pathogen in immunocompromised patients.     

复发、转移子宫颈癌调强放疗和三维适形放疗的比较

 目的　比较调强放疗（IMRT）和三维适形放疗（3DCRT）治疗复发、转移子宫颈癌的疗效、剂量学及毒副作用。方法　回顾性分析治疗后复发转移子宫颈癌62例,其中IMRT组29例,3DCRT组33例,均行直线加速器6 MV X线放疗,单次剂量1.8～2.2 Gy,每周5次,共18～33次,处方剂量40～60 Gy,中位剂量52.8 Gy。同时对IMRT组的患者设计行3DCRT,给予相同的处方剂量,比较危及器官（OAR）受照射剂量。结果　IMRT组膀胱和小肠的最高剂量分别为（4642.71±805.53）cGy和（4240.36±572.51）cGy,低于3DCRT组的（5057.53±1998.03） cGy和（5953.99±1180.81）cGy（P＜0.05);IMRT计划中PTV的最高剂量（5245.68±365.26）cGy高于3DCRT的最高剂量（4801.27±346.25）cGy,差异具有统计学意义（P＜0.05）。IMRT组1、2、3年生存率分别为65.5 %(19／29)、42.1 %(8／19)、25.0 %（2／8）,中位生存时间为19个月,28例死亡病例中,21例死于肿瘤进展,7例死于远处转移;3DCRT组：1、2、3年生存率分别为60.6 %（20／33）、35.0 %（7／20）、14.3 %(1／7),中位生存时间为17个月,32例死亡病例中,24例死于肿瘤进展,8例死于远处转移。IMRT组和3DCRT组比较,1、2、3年生存率差异无统计学意义（均P＞0.05）。IMRT组的不良反应的发生率明显低于3DCRT组,尤其是Ⅰ级和Ⅱ级,IMRT组为24.1 %（7／29）,3DCRT组为33.5 %（11／33）。结论　IMRI对于复发转移子宫颈癌疗效较3DCRT更好,可以在提高肿瘤剂量的同时减少正常组织的受照体积和剂量,减少不良反应的发生。     

Abrogation of Microsatellite-instable Tumors Using a Highly Selective Suicide Gene/Prodrug Combination

A substantial fraction of sporadic and inherited colorectal and endometrial cancers in humans is deficient in DNA mismatch repair (MMR). These cancers are characterized by length alterations in ubiquitous simple sequence repeats, a phenotype called microsatellite instability. Here we have exploited this phenotype by developing a novel approach for the highly selective gene therapy of MMR-deficient tumors. To achieve this selectivity, we mutated the VP22FCU1 suicide gene by inserting an out-of-frame microsatellite within its coding region. We show that in a significant fraction of microsatellite-instable (MSI) cells carrying the mutated suicide gene, full-length protein becomes expressed within a few cell doublings, presumably resulting from a reverting frameshift within the inserted microsatellite. Treatment of these cells with the innocuous prodrug 5-fluorocytosine (5-FC) induces strong cytotoxicity and we demonstrate that this owes to multiple bystander effects conferred by the suicide gene/prodrug combination. In a mouse model, MMR-deficient tumors that contained the out-of-frame VP22FCU1 gene displayed strong remission after treatment with 5-FC, without any obvious adverse systemic effects to the mouse. By virtue of its high selectivity and potency, this conditional enzyme/prodrug combination may hold promise for the treatment or prevention of MMR-deficient cancer in humans.     

婴幼儿病毒性肺炎的病毒病原学及临床特点

[目的]探讨本地区引起小儿病毒性肺炎的病原学概况及其相应临床表现,为临床诊治提供依据.[方法]应用ELISA法检测325例婴幼儿肺炎患儿急性期血清中五种常见病毒（RSV、IFV、CMV、ADV、PIV）血清特异性IgM;用速率散射比浊法测定C-反应蛋白（CRP）和免疫球蛋白.用碱性磷酸酶标记链霉卵白素（S-A/AP）法检测人外周血T淋巴细胞亚群.[结果]325例婴幼儿肺炎中共检出病毒感染157例,病毒感染率48.3%,单一病毒感染113例（72.0%）,其中合胞病毒（RSV）32株（20.4%）,流感病毒（IFV）27株（17.2%）,腺病毒（ADV）22株（14.0%）,副流感病毒（PIV）17株（10.8%）,巨细胞病毒（CMV）15株（9.6%）;混和病毒感染共44例（28.0%）,以IFV＋PIV 15例（9.6%）多见.RSV肺炎患儿喘息重,缺氧征明显;ADV肺炎患儿以高热和中毒表现为主;IFV和PIV肺炎患儿主要表现为咳嗽重、喘息轻;CMV肺炎患儿除一般症状外,常伴肝功能异常.1～3个月、～12月、～3岁三个年龄组病毒性肺炎患儿血清lgG、lgA、lgM与对照组比较,差异均无显著性（P＞0.05）;而T细胞亚群与对照组比较,CD3、CD4、CD4/CD8均降低、CD8升高,差异均有显著性（P＜0.05）.[结论]病毒感染是婴幼儿肺炎的主要病原,各类病毒性肺炎各有其临床特点,细胞免疫功能紊乱在小儿病毒性肺炎发病机制中起重要作用.     

临朐县1999～2003年公共场所从业人员HBsAg检测分析

[目的]了解公共场所从业人员HBsAg携带状况，加强监督力度。[方法]对临朐县1999～2003年公共场所从业人员4711人进行HBsAg检测情况进行分析。[结果]检测公共场所从业人员4711人，HBsAg阳性155人，阳性率为3．29％；不同年份阳性率的差异无统计学意义（P〉0．05）；不同性别、各年龄组阳性率的差异有统计学意义（P〈0．01）。[结论]临朐县公共场所从业人员HBsAg携带状况基本保持不变，应引起重视防止疾病的传播。     

2004年住院患者抗菌药物使用情况调查分析

目的：了解我院抗菌药物应用的合理性。方法：抽取2004年我院住院患者的所有处方（电子处方），统计抗菌药物的应用品种、数量、消耗金额及使用频度，结合病历分析我院应用抗菌药物的合理性。结果：β-内酰胺类抗生素是我院抗感染的首选用药。结论：我院抗感染用药以疗效可靠、不良反应小、价格便宜的抗生素为主。     

自身免疫性甲状腺疾病实验室诊断

目的：采用TPOAb、TGb、FT3、FT4五项指标对自身免疫性甲状腺疾病（AITD）提供诊断和鉴别诊断的实验室依据。方法：采用放射免疫（受体）分析法测量165例AITD患者血清中TPOAb、TGAb、TRAb、FT3、FT4水平。结果：桥本甲状腺炎组TPOAb和TGAb升高与Graves病组、桥本甲亢组比较差异有显著性（P〈0．001）；TRAb、FT3、FT4正常与Graves病组、桥本甲亢组结果差异有显著性（P〈0．001）；Graves组和桥本甲亢组TRAb、FT3、FT4升高差异无显著性（P〉0．05），而TPOAb和TGAb结果升高差异有显著性（P〈0．001）。结论：上述五项实验室指标对AITD诊断、鉴别诊断及治疗原则、方法的选择具有重要意义。     

Role of macrophage colony-stimulating factor in the differentiation and expansion of monocytes and dendritic cells from CD34+ progenitor cells

The present study focused on whether it is possible to expand monocytic cells from CD34+ progenitor cells by using macrophage colony-stimulating factor (M-CSF) in the absence and presence of mast cell growth factor (MGF) and IL-6. It was demonstrated that CD34+ cells differentiate without expansion to functional mature monocytic cells in the presence of M-CSF or combinations of M-CSF plus IL-6 and MGF. A different response pattern was observed for the number of clonogenic cells. The addition of IL-6 or both IL-6 and MGF to M-CSF containing cultures resulted in significant higher numbers of colony-forming unit-macrophage (CFU-M) as tested in clonogenic and 3H-thymidine assays. Furthermore, M-CSF plus both IL-6 and MGF appeared to be the most potent combination to preserve the monocytic precursor in cell suspension culture assays. These results indicate that IL-6 and MGF in conjunction with M-CSF affect CD34+ cells especially at precursor level without distinct effect on the more mature stages. Secondly we studied whether M-CSF is only critical for the monocytic lineage or also affects dendritic cell (DC) development. Indeed, we were able to culture CD83+ DC from CD34+ progenitor cells in the presence of M-CSF in conjunction with TNF-alpha, IL-4, and MGF although their absolute number is almost threefold lower than the number of CD83+ cells yielded from GM-CSF plus TNF-alpha, IL-4, and MGF stimulated CD34+ cells.     

Safety and pharmacokinetics of multiple doses of recombinant human CuZn superoxide dismutase administered intratracheally to premature neonates with respiratory distress syndrome

OBJECTIVES: To examine the safety and pharmacokinetics of multiple intratracheal (IT) doses of recombinant human CuZn superoxide dismutase (rhSOD) in premature infants with respiratory distress syndrome who are at risk for developing bronchopulmonary dysplasia (BPD). Methods. Thirty-three infants (700 to 1300 g) were randomized and blindly received saline, 2.5 mg/kg or 5 mg/kg rhSOD IT within 2 hours of surfactant administration. Infants were treated every 48 hours (as long as endotracheal intubation was required) up to 7 doses. Serial blood and urine studies, chest radiographs, neurosonograms, SOD concentration and activity measurements, and tracheal aspirate (TA) inflammatory markers were assessed throughout the 28-day study. RESULTS: SOD concentrations in serum (0.1 [0.05/0.15] microg/mL-geometric mean with lower/upper confidence intervals), tracheal aspirates (TA) (0.2 [0.1/0.3] microg/mL) and urine (0.3 [0.2/0.4] microg/mL) were similar at baseline in all 3 groups and did not change significantly in the placebo group. In the rhSOD treatment groups, SOD concentrations were increased on day 3 and did not change significantly thereafter over the 14-day dosing period (also measured on days 5, 7, and 13). SOD concentrations averaged 0.4 [0.3/0.5] microg/mL in serum, 0.8 [0.6/1.2] microg/mL in TA and 1.1 [1.0/1.3] microg/mL in urine for the low-dose group and 0.6 [0.5/0.7] microg/mL in serum, 1.1 [0.9/1.5] microg/mL in TA, and 2.2 [1.6/2.9] microg/mL in urine for the high-dose group over the 14-day dosing period. Enzyme activity directly correlated with SOD concentration and rhSOD was active even when excreted in urine. TA markers of acute lung injury (neutrophil chemotactic activity, albumin concentration) were lower in the rhSOD agroups compared with placebo. No significant differences in any clinical outcome variable were noted between groups. CONCLUSIONS: These data indicate that multiple IT doses of rhSOD increase the concentration and activity of the enzyme in serum, TA and urine, reduce TA lung injury markers and are well-tolerated. Further clinical trials examining the efficacy of rhSOD in the prevention of BPD are warranted.