Transplantation of enriched and purged peripheral blood progenitor cells from a single apheresis product in patients with non-Hodgkin's lymphoma

High-dose chemotherapy with or without radiotherapy followed by autologous transplantation of hematopoietic progenitor cells is an effective treatment for patients with high-risk or relapsed non- Hodgkin's lymphoma. Chemotherapy and/or hematopoietic growth factors have been used to mobilize progenitor cells in the peripheral blood for transplantation. However, the mobilized blood cell products have been found to be frequently contaminated with tumor cells, and techniques have not been developed to purge tumor cells from these products. In addition, the minimum number of hematopoietic progenitor cells required for engraftment has not yet been fully elucidated. We treated 21 patients with a single infusion of cyclophosphamide (4 g/m2) followed by daily administration of granulocyte colony-stimulating factor (G- CSF). After recovery of the white blood cell count, a single 3-hour apheresis collection was performed. The apheresis product was then applied to a discontinuous Percoll gradient. The low-density fractions resulting from this separation procedure were enriched for CD34+ progenitor cells (total cell yield, 19.5%; CD34+ cell recovery, 81.2%). These enriched cellular products were treated with a panel of anti-B cell or anti-T cell monoclonal antibodies and complement in an effort to remove residual tumor cells. After treatment of the patient with myeloablative therapies, the enriched and purged cells were reinfused. Hematologic recovery was rapid, with median neutrophil engraftment in 10 days [absolute neutrophil count (ANC), greater than 0.5 x 10(9)/L] and 11 days (ANC, greater than 1.0 x 10(9)/L). Median platelet transfusion independence required 13 days. The rapidity of multilineage engraftment correlated with the number of CD34+ cells per kilogram that were infused. Patients who received more than 2 x 10(6) CD34+ cells per kilogram had rapid hematologic engraftment, whereas those patients transplanted with less than 2 x 10(6) CD34+ cells per kilogram had slower platelet recovery. Modeling studies using a lymphoma cell line with a t(14; 18) chromosomal translocation demonstrated the successful removal of tumor cells assayed using the polymerase chain reaction (PCR) after the processing and purging. Four of the 21 patients had PCR- detectable lymphoma cells in the bone marrow and peripheral blood; however, the enriched and purged blood products reinfused in all four did not contain detectable tumor cells.(ABSTRACT TRUNCATED AT 400 WORDS)     

前路与后路腰椎间盘切除后Cage植入治疗与射频治疗改善椎间盘源性下腰痛的效果比较

目的:比较采用射频疗法与前路和后路椎体间融合术治疗椎间盘源性下腰痛疗效差别。方法:①于1999-01/2005-08在解放军总医院骨科、北京大学第三医院骨科有完整临床资料的椎间盘源性腰痛患者53例,男33例,女20例;年龄33～58岁。纳入对象影像学检查显示无明显神经根受压、腰椎不稳等其他腰椎疾病;符合国际疼痛学会1988年制定的椎间盘造影阳性标准;均对治疗方案知情同意。②所有入选病例依据治疗方法不同分为3组:射频组21例:行经皮穿刺射频汽化成型术;前路椎体间融合组14例:行腹膜外椎间盘切除、Cage植入、结合后路经椎板关节突螺钉固定、髂骨取骨植骨融合术;后路椎体间融合组18例:行后路椎间盘切除、椎间Cage植入结合椎弓根螺钉内固定后外侧植骨融合术。③统计手术时间和出血量,术后1年进行放射学评估融合情况。采用目测类比评分评估患者疼痛程度(无痛为0分,腰痛到无法忍受的最大程度为10分)。参照Oswestry功能障碍指数评估患者腰部功能恢复程度(分值越高表示功能受限程度越重)。分别于术前、术后1年进行评分。改善率=[(术前评分-术后评分)/术前评分]×100%。分级标准,优:改善率≥75%。良:改善率50%～75%;可:改善率25%～50%;差:改善率<25%。④计量资料差异比较采用连续型重复测量资料的方差分析。结果:椎间盘性下腰痛患者53例均进入结果分析。①手术时间和出血量比较:后路椎体间融合组手术时间和手术出血量明显长于/高于其他2组(P<0.05),射频组明显短于/小于前路椎体间融合组(P<0.05)。②术后疼痛改善情况:射频组良9例,可10例,差2例;前路椎体间融合组和后路椎体间融合组疼痛评分改善均为优。射频组目测类比评分改善率明显低于其他2组(P<0.05)。③术后功能改善情况:射频组良5例,可13例,差3例;前路椎体间融合组优12例,良2例;后路椎体间融合组优6例,良12例。射频组Oswestry功能障碍指数改善率明显低于前、后路椎体间融合组(P<0.05),前路椎体间融合组Oswestry功能障碍指数改善率明显高于后路椎体间融合组(P<0.05)。④椎体融合情况:前、后路椎体间融合组术后1年椎体融合率均为100%。结论:射频治疗创伤小但疗效差,前路椎间盘切除Cage植入结合后路经椎板关节突螺钉固定与后路椎间盘切除椎间融合结合椎弓根固定可明显改善疼痛,前者创伤相对小,功能恢复更好,是治疗椎间盘源性腰痛的最佳选择。     

An improved clonal excess assay using flow cytometry and B-cell gating

In humans with B-cell malignancies, the presence of monoclonal B lymphocytes (clonal proliferation) can be detected by comparing the fluorescence intensity distributions of lymphocytes stained with anti- kappa and anti-lambda reagents. The sensitivity of previously described single-color immunofluorescence techniques to low levels of clonal excess is limited by background from cytophilic immunoglobulins on non- B cells and by the low proportion of circulating B cells in individuals with minimal disease. We have used two-color immunofluorescence and B- cell gating to develop an improved assay that avoids false positives due to non-B cells, without requiring restrictive light scatter gates that may exclude true positives. This method is sensitive to 0.2% monoclonal B cells admixed with fresh normal lymphocytes, to 0.6% monoclonal B cells admixed with normal lymphocytes that have been stored for up to 72 hours, and readily detects 1% monoclonal cells in patient specimens. The two color B-cell gated assay offers sensitivity equivalent to the single-color assay and improved specificity for detection of low levels of clonal excess.     

TNF-α值与2型糖尿病胰岛素抵抗的关系

目的　探讨肿瘤坏死因子 -α(TNF -α)与 2型糖尿病胰岛素的关系。方法　用homa -R法选取有胰岛素抵抗 (IR组 )和无胰岛素抵抗 (NIR组 )的 2型糖尿病患者各 2 0例。另以 2 0名年龄及体重指数相匹配的人群的作为对照组 ,分别检测其空腹血糖、胰岛素 ,计算Homa -R值 ,用ELISA法检测TNF -α。结果　IR组的TNF -α与NIR组及对照组相比较明显增高 ,有统计学意义(P <0 0 5 ) ,IR组的空腹胰岛素水平及胰岛素抵抗指标Homa -R也显著高于NIR组及对照组 (P值为 <0 0 1) ,而三组间年龄、BMI、血脂、脂蛋白、血压均无显著性差异 (P >0 0 5 )。结论　血浆TNF -α水平与胰岛素抵抗呈密切相关 ,在 2型DM中有重要病理意义 ,可作为评估有无胰岛素抵抗的一项重要参考指标     

Chronic low back pain in primary care: a prospective study on the management and course

BACKGROUND: There is little evidence about the management and course of chronic low back pain in primary care. OBJECTIVES: Our aim was to describe the course of chronic low back pain and the performed diagnostic and therapeutic procedures for patients with chronic low back pain in general practice. METHODS: Twenty-six GPs involved in the Registration Network Family Practices participated in this prospective follow-up study. All patients and GPs were asked to complete questionnaires at baseline and at 4, 8 and 12 months follow-up. RESULTS: The GPs provided information about diagnostic and therapeutic procedures concerning 524 patients with chronic low back pain. Diagnostic tests other than history-taking and physical examination were not frequently used. Medication, mostly NSAIDs, was the most frequently used type of treatment (21.6%). The most frequent referrals concerned physiotherapy (16.3%) and neurology or neurologic surgery (6.3%). Information about the course of their chronic low back pain was provided by 368 patients participating in our study. The course of chronic low back pain appeared to be quite stable, as there was only a slight improvement in pain intensity and physical functioning over the 12 months of follow-up. CONCLUSIONS: A variety of options for the treatment and referral of chronic low back pain patients is available for and used by GPs. Efforts should be made to establish which diagnostic and therapeutic procedures are the most effective for chronic low back pain.      

Hormone-induced refractoriness to mammary carcinogenesis in Wistar- Furth rats

One of the most consistent results in the epidemiology of human breast cancer is the inverse relationship of risk and early full-term parity. The goal of this study was to investigate the molecular mechanisms through which early full-term pregnancy protects the breast from cancer development. We used Wistar-Furth (WF) rats as our experimental system and mimicked pregnancy using estrogen and progesterone (E/P). Sexually mature female rats were treated with steroid hormones for 21 days and after 28 days of gland involution, the rats were administered MNU. Rats that received a high dose of 20 microg E and 20 mg P exhibited an 82% reduction in the incidence of mammary adenocarcinomas as compared to the rats receiving only blank pellets. Decreasing doses of E/P were partially protective suggesting that complete differentiation of the gland was not required for refractoriness. We measured the RNA expression levels of several target genes involved in the regulation of mammary cell proliferation and/or differentiation including estrogen receptor (ER) and progesterone receptor (PR), cyclins D1 and D2, the cell cycle inhibitors p16, p21 and p27, and the tumor suppressor p53. At the time of MNU treatment we found no significant differences in the expression of these genes, with the possible exception of p21, indicating that hormone treatment did not result in constitutive changes in expression levels. The numbers of apoptotic cells were low and comparable in the hormone exposed and age-matched virgin gland (AMV) at the time of carcinogen challenge and remained low for 8 days after MNU treatment. The number of BrdU-labeled cells at the time of carcinogen challenge were also low in both the AMV (1.8%) and hormone exposed (0.8%) animals. In contrast, cell proliferation in the AMV (5.7%) was significantly different from both the parous involuted (1.2%) and the E/P-treated involuted (1.5%) animals 8 days after MNU treatment. We interpret these data to indicate that hormone treatment results in mammary epithelial cells that have persistent alterations in intracellular pathways governing proliferation responses to carcinogens.