Basic fibroblast growth factor expression in human bone marrow and peripheral blood cells

We have shown previously that basic fibroblast growth factor (bFGF) is a mitogen for human bone marrow (BM) stromal cells and that bFGF stimulates myelopoiesis in primary BM cultures. In this article, we demonstrate the presence of bFGF in two cell lineages in human BM and peripheral blood as well as the deposition of bFGF into the extracellular matrix of BM stromal cell cultures. In immunofluorescence experiments on BM and peripheral blood smears, megakaryocytes and platelets stained strongly for bFGF, whereas weaker staining was observed in immature and mature cells of the granulocyte series. The presence of bFGF in platelets was confirmed by enzyme-linked immunosorbent assay as well as by immunoprecipitation followed by immunoblotting. bFGF was synthesized by BM stromal cell cultures and was found either cell associated or localized in the nucleus and the nucleoli, and its location was dependent on the fixation procedure used. Addition of exogenous bFGF to stromal cells showed the presence of extracellular binding molecules for this cytokine. bFGF could be released from these sites by soluble heparin or phosphatidylinositol- specific phospholipase C. This study supports the role of bFGF as a stromal cell mitogen and stimulator of myelopoiesis. The data indicate that the stromal cells produce bFGF and that their extracellular matrix can serve as a reservoir for this growth factor. In addition, the results suggest a possible involvement of bFGF in platelet function as well as in megakaryocytopoiesis.     

Interaction of single-chain urokinase with its receptor induces the appearance and disappearance of binding epitopes within the resultant complex for other cell surface proteins

Binding of urokinase-type plasminogen activator (uPA) to its glycosylphosphatidylinositol-anchored receptor (uPAR) initiates signal transduction, adhesion, and migration in certain cell types. To determine whether some of these activities may be mediated by associations between the uPA/uPAR complex and other cell surface proteins, we studied the binding of complexes composed of recombinant, soluble uPA receptor (suPAR) and single chain uPA (scuPA) to a cell line (LM-TK- fibroblasts) that does not express glycosylphosphatidylinositol (GPI)-anchored proteins to eliminate potential competition by endogenous uPA receptors. scuPA induced the binding of suPAR to LM-TK- cells. Binding of labeled suPAR/scuPA was inhibited by unlabeled complex, but not by scuPA or suPAR added separately, indicating cellular binding sites had been formed that are not present in either component. Binding of the complex was inhibited by low molecular weight uPA (LMW-uPA) indicating exposure of an epitope found normally in the isolated B chain of two chain uPA (tcuPA), but hidden in soluble scuPA. Binding of LMW-uPA was independent of its catalytic site and was associated with retention of its enzymatic activity. Additional cell binding epitopes were generated within suPAR itself by the aminoterminal fragment of scuPA, which itself does not bind to LM-TK- cells. When scuPA bound to suPAR, a binding site for alpha 2-macroglobulin receptor/LDL receptor-related protein (alpha 2 MR/LRP) was lost, while binding sites for cell-associated vitronectin and thrombospondin were induced. In accord with this, the internalization and degradation of cell-associated tcuPA and tcuPA-PAI- 1 complexes proceeded less efficiently in the presence of suPAR. Further, little degradation of suPAR was detected, suggesting that cell- bound complex dissociated during the initial stages of endocytosis. Thus, the interaction of scuPA with its receptor causes multiple functional changes within the complex including the dis-appearance of an epitope in scuPA involved in its clearance from the cell surface and the generation of novel epitopes that promote its binding to proteins involved in cell adhesion and signal transduction.     

Anti-Ata causing mild hemolytic disease of the newborn

We report here a case of moderately severe hemolytic disease of the newborn (HDN) due to anti-Ata. The gravida 5 proposita was group A rr and previously was found to have anti-Ata and -D. At the 35-week mark of this pregnancy, her anti-Ata demonstrated a titer of 256, score 79. Fluid obtained by amniocentesis at 36 weeks showed an optical density at 450 nm of 0.08 (midzone). The baby was delivered at 38 weeks by cesarean section. The cord cells were group A rr with a 3+ direct antiglobulin test. The dichloromethane eluate of the infant's cells demonstrated anti-Ata specificity only. At birth, the infant's total bilirubin (TB) was 2.1 mg per dl and the hematocrit level (Hct) was 33.8 percent. Within 8 hours, the TB had risen to 3.8 mg per dl. Phototherapy was initiated at 7-1/2 hours and maintained for 40 hours. The infant's TB rose to a maximum level of 10.5 mg per dl 24 hours after phototherapy was discontinued. At discharge 4 days postpartum, the infant's TB had dropped to 9.2 mg per dl, and the Hct value was 38 percent. On a visit 7 days postpartum, the infant's TB level had fallen to 6.5 mg per dl and the hct value was 38 percent. Transfusions were not necessary.     

人工髓核假体置入治疗腰椎间盘突出症的疗效分析

目的：观察已在临床初步开展起来的人工髓核置换术治疗腰椎间盘突出症的中、远期临床疗效及并发症,分析其对策。 方法：纳入2002-02/2004-08南方医科大学附属南方医院脊柱骨病外科采用单枚人工髓核假体置换术治疗单节段腰椎间盘突出症患者98例,获得24～48个月随访患者75例,按平均随访时间达24,36,48个月,分为24个月组（n=30）,36个月组（n=23）,48个月组（n=22）。选同期采用单纯椎间盘髓核摘除术患者30例作为对照组,评估各组术后临床疗效,主观症状采用Oswestry功能障碍指数问卷表（0%表示正常,越接近100%则功能障碍越严重）和Prolo功能评分表（小于或等于5分为差,6～7分为中等,8～10为优）评价,分析术后影像学检查并测量手术节段活动度和椎间隙高度变化情况,同时观察假体位置情况,腰椎MRI观察假体位置和软骨终板的信号变化情况。腰椎活动度=（腰椎中立角度-前屈角度）＋（后伸角度-腰椎中立角度）=后伸角度-前屈角度;为消除X射线放大率的影响,椎间隙高度变化情况采用病变椎间隙后缘高度与上位椎体中部矢状径的比值表示。 结果：75例获得24～48个月随访者,全部进入结果分析。①48个月组2例、36个月组1例发生假体脱出,二次手术取出。其余患者术后临床症状均明显缓解,疼痛消失。②24,36,48个月组及对照组术后末次Oswestry功能障碍指数均较术前降低,差异有显著性意义（14.2%,52.1%;15.5%,55.2%;15.1%,53.6%;15.5%,51.5%;P〈0.05）。③24,36,48个月组及对照组术后末次Prolo能评分均较术前升高,差异有显著性意义（8.5,4.6分;8.6,4.5分;8.7,4.3分;8.4,4.2分;P〈0.05）。④24,36,48个月组同期手术节段腰椎活动度均高于对照组,差异有显著意义（P〈0.05）。⑤24个月组手术节段椎间高度较术前降低约4%、36个月组降低约12%、48个月组降低约18%、对照组较术前降低约25%,各组术前和术后椎间隙高度比值比较,差异具有显著性意义（P〈0.05）。⑥主要并发症：早期出现术后一过性腰痛24例,假体脱出3例。中、远期发现假体下沉32例,软骨终板损伤39例。 结论：单枚人工髓核假体置换治疗腰椎间盘突出症中、远期随访临床疗效肯定,但存在较严重并发症,应慎重开展此项手术。     

自主研制镍钛记忆合金左心耳封堵器封闭左心耳对实验动物左心功能的影响

目的:应用经胸彩色多普勒超声技术评价自主研制的镍钛记忆合金左心耳封堵器封闭左心耳对实验动物猪左心房、左心室功能的影响。方法:实验于2005-09/2006-08在南京医科大学第一附属医院江苏省实验动物中心完成。①实验分组:选用苏钟小型种猪17只,随机分为实验组12只和对照组5只。②实验干预:实验组12只苏钟小型种猪使用自主研制的左心耳封堵器(发明专利号码:200610037789.3,公开号CN1799521,由镍钛合金骨架、多聚四氟乙烯膜和传送连接部分等构成。其外观呈单盘状,封堵器的左心房面呈圆盘状,直接连接放入心耳内的圆柱体结构)行左心耳封堵,对照组5只手术步骤相同而不采用封堵器行左心耳封堵。③实验评估:两组动物分别于术前、术后1周、2周、4周采用经胸超声心动图检查观察心功能的改变,测量左心房内径、最大及最小容积、左房射血分数、左心房搏出量、血流分数等左房功能参数以及左室射血分数、左室短轴缩短率、Tei指数、E/A比值等指标。结果:①实验动物数量分析:在施行左心耳封堵后,1头猪于术中出血过多并出现室颤后死亡,1头猪因封堵器脱入左房,卡在二尖瓣口导致死亡。其余动物封堵效果良好。②两组动物术后1,2,4周左房功能指标各参数与术前比较无明显变化(P>0.05);与术前相比,实验组术后1周、2周左室射血分数、左心室短轴缩短率、E/A比值分别由术前的0.70±0.04、0.39±0.03、1.33±0.28降低至术后1周的0.59±0.05、0.31±0.03、0.95±0.11(P<均0.01)及术后2周的0.62±0.05、0.33±0.05、0.90±0.05(P<均0.01);Tei指数由术前的0.48±0.02增加至术后1周的0.59±0.03(P<0.01)及术后2周的0.58±0.04(P<0.01)。对照组手术前后左室功能指标差异无显著性。结论:自主研制左心耳封堵器可以有效的封堵左心耳;左心耳封堵后短期内对实验动物左房功能无明显影响;封堵后短期内对左心室功能具有短期的减弱,更长期的安全性有待于进一步研究。     

Donation reactions among autologous donors

Studies of risk factors associated with reactions among autologous blood donors have been limited. Therefore, 2091 autologous and 4737 homologous donations were examined. Donors at greatest risk for reaction were autologous donors who had reactions at first donation; among 45 who made repeat donations for the same surgery, 17 (38%) had repeat reactions. The group least likely to experience reactions were the autologous donors greater than or equal to 66 years old; they experienced a 1.9 greater than or equal to percent (6/310) incidence of reactions. More reactions were seen in both autologous and homologous donors in the categories of first-time donor, female gender, decreasing age, and lower weight. Multiple logistic regression analysis showed that all of these variables were independent predictors of donor reaction, with first-time donation (odds ratio, 2.4) and female gender (odds ratio, 1.9) being the strongest predictors of reaction. Donor room personnel should be alerted that autologous donors who react at first donation are very likely to react at subsequent donations. Contrary to common concern, elderly autologous donors are least likely to have reactions.     

T lymphocytes specific for immunoglobulin allotype. I. Igh- 1(b)-specific T cells demonstrated by suppression in vivo and cytotoxicity in vitro

We show that determinants of IgG(2a) of C57BL/6 mice (Igh-1(b)) stimulate allotypespecific T cells in BALB/c mice. Such cells are detected in two different functional assays; chronic allotype suppression and T cell-mediated cytotoxicity. A population of suppressor T cells capable of inducing chronic Igh-1(b) suppression was demonstrated by rosetting procedures to possess Igh-1(b)-specific receptors, a result interpreted as indicating that suppressor T cells may act directly upon allotype-bearing B cells. From similar populations we were also able to demonstrate Igh-1(b)-specific cytotoxic T cells. Such cells were lytic for target myeloma cells expressing the Igh-1(b) allotype of IgG28, and were ineffective against a variant cell line failing to express Igh-1(b), and other target cell lines expressing different allotypes or isotypes. The similar specificity of suppressor T cells and cytotoxic T lymphocytes for Igh-1(b) allotype raises the possibility that the target in allotype suppression is a B cell, and that allotype-specific cytotoxic T cells may play some role in regulation of allotype expression in the suppressed state.     

Low risk of viral infection after administration of vapor-heated factor VIII concentrate. International Investigator Group

A multicenter prospective study was carried out to evaluate whether a vapor-heated factor VIII concentrate transmitted blood-borne viral infections over a surveillance period of 15 months. Thirty-five patients with hemophilia and von Willebrand disease who had never received any blood components were treated. Twenty-eight were analyzed and found not to have non-A, non-B hepatitis. Sera from 20 of these 28 patients were also tested for the antibody to the hepatitis C virus. None had sero-converted during the follow-up period. None of the patients analyzed developed markers of the hepatitis B virus (n = 17) or the human immunodeficiency virus (n = 31). This vapor-heated factor VIII concentrate carries a low risk of transmitting hepatitis and human immunodeficiency virus infection.     

高效液相色谱法对四种双氯灭痛软膏剂透皮效果的研究

目的：本实验采用高效液相色谱法,以离体小白鼠腹部皮肤为透皮屏障,比较了四种双氯灭痛软膏剂的透皮效果,方法：实验用法国Gilson公司的HPLC仪,以C18为固定相,甲醇－水（含HAc 0.36%)=80:20为流动相,紫外检测波长280nm。平均回收率为99．59％,RSD＜3．28％,检测限为1．0ug/ml。结果：1号样品透皮率最高,3号与1号相比较透皮吸收无显著性差异（P＞0．05）,2号或4号分别与1号相比,其透皮吸收均有显著性差异（P＜0．05）,结论：1号品值得进一步开发。