透析用水及透析液中内毒素污染状况的分析

目的　研究透析用水、透析液内毒素及细菌污染状况。方法　对北京１８ 家医院透析室反渗水、透析液用改良鲎试验检测内毒素,用血琼脂培养基做细菌培养,用ＥＬＩＳＡ法测定患者血浆白介素（ＩＬ）１、ＩＬ６ 及肿瘤坏死因子（ＴＮＦ）α。结果　反渗水内毒素为（０．１１６±０ ．０６３） ＥＵ／ｍｌ;细菌培养有两家医院阳性,均为１００ ＣＦＵ／ｍｌ。Ｂ浓缩透析液内毒素为（０．４６ ±０．３５） ＥＵ／ｍｌ,而Ａ浓缩透析液内毒素仅（０．０９１±０ ．０８４） ＥＵ／ｍｌ（Ｐ＜０ ．００１） ;Ｂ浓缩透析液细菌培养１１ 家阳性,其中８ 家≥２ ０００ ＣＦＵ／ｍｌ,Ａ浓缩透析液细菌培养阳性仅３ 家,均＜ ２ ０００ ＣＦＵ／ｍｌ。１６ 例患者中有５ 例透析器入口透析液内毒素＞０．５ ＥＵ／ｍｌ,且透后血ＩＬ６ 及ＴＮＦα显著增高。结论　目前透析用水及透析液存在内毒素及细菌的污染,定期检测反渗水、透析液的内毒素水平及进行细菌培养,定期消毒反渗水装置及透析液容器,对减少热源反应至关重要。     

慢性肾衰患者血清及尿液γ-干扰素的检测及其意义

采用ＥＬＩＳＡ双抗体夹心法检测７１例慢性肾衰（ＣＲＦ）患者及４０例健康成人血及尿的γ－干扰素（ＩＦＮ－γ）水平。结果显示：ＣＲＦ患者血清及尿液ＩＦＮ－γ含量分别为（６２６２±２．６８）ｎｇ／Ｌ和（４３．０１±２．５７）ｎｇ／Ｌ，显著低于正常对照组，［（１０５±７．１０）ｎｇ／Ｌ和（７５±６．１４）ｎｇ／Ｌ］（Ｐ＜０００１），但与尿素氮（ＢＵＮ）及血清肌酐（Ｓｃｒ）无显著相关性（血清及尿ＩＦＮ－γ与ＢＵＮ、Ｓｃｒ相关系数分别是ｒ＝０．２４２５、０３０５６、０１５３３和０１７５０，Ｐ＞００５）。结论：ＩＦＮ－γ作为一种细胞因子参与肾脏病发生、发展过程中的网络调节，血清及尿液中ＩＦＮ－γ水平受到机体Ｔ淋巴细胞及ＮＫ细胞数量及功能影响。     

消炎痛雾化吸入对慢性支气管炎的疗效观察

采用随机、单盲、安慰剂及正常对照的方法，观察了４８例慢性支气管炎（慢支）急性发作期患者痰及血中６－酮－前列腺素Ｆ１ａ（６－ｋｅｔｏ－ＰＧＦ１ａ）、血栓素Ｂ２（ＴＸＢ２）含量的变化以及它们与疾量，痰干／湿比，肺活量实测值占预计值百分比（ＶＣ％），１秒用力肺活量占用力肺活量百分比（ＦＥＶ１％）的相关性。并观察了消炎痛对它们的影响。结果表明：慢支患者痰及血中６－ｋｅｔｏ－ＰＧＦ１ｕ及ＴＸＢ２均高于正常对照组（Ｐ＜０．０１）。痰及血中６—ｋｅｔｏ－ＰＧＦ１ａ和ＴＸＢ２与痰量和疾干／湿比里正相关（Ｐ＜０．０１）；与ＶＣ％和ＦＥＶ１％呈负相关（Ｐ＜０．０５，Ｐ＜０．０１）。与安慰剂组相比消炎痛雾化吸入可明显降低慢支患者的痰量，痰粘度和血沉（Ｐ＜０．０１）；明显提高ＶＣ％和ＦＥＶ１％（Ｐ值分别小于０．０５，Ｐ＜０．０１）；明显缩短病程（Ｐ＜０．０５）。     

健康老年人骨髓红系和粒—单系祖细胞的体外培养研究

采用造血祖细胞培养技术研究了健康老年人骨髓红系和粒－单系祖细胞的增殖能力及骨髓红系和粒－单系祖细胞分别对红细胞生成素（ＥＰＯ）和粒－单系集落刺激因子（ＧＭ－ＣＳＦ）的反应能力。发现健康老年人红系爆式集落形成单位（ＢＦＵ－Ｅ）、红系集落形成单位（ＣＦＵ－Ｅ）和粒－单系集落形成单位（ＣＦＵ－ＧＭ）的集落数比健康非老年人明显减少；ＢＦＵ－Ｅ和ＣＦＵ－Ｅ对ＥＰＯ以及ＣＦＵ－ＧＭ对ＧＭ－ＣＳＦ的反应曲线低平     

慢性肾功能不全患者血清可溶性Fas及相关因子的变化

目的 研究慢性肾功能不全患者血清可溶性Ｆａｓ／ＡＰＯ－（ｓＦａｓ）及相关因子的变化。方法 采用ＥＬＩＳＡ法检测２０名正常人和６０例慢性肾功能不全患者血清ｓＦａｓ及ＩＬ－６、ＴＮＦ－α水平。结果 肾功能代偿期ｓＦａｓ、ＩＬ－６和ＴＮＦ－α无明显改变;肾功能代偿期ｓＦａｓ开始增高,其升高与血甭ＢＵＮ和Ｃｒ明显相关。ＩＬ－６及ＴＮＦα在肾功能失代偿期明显变化,进入肾功能衰竭期后显著升高。其中ＩＬ－６ 关     

rhIL—6,rhGM—CSF和rhEPO对人造血干细胞的作用

重组人白细胞介素 ６和重组人粒－单细胞集落刺激因子与正常人造血干细胞培养１周后，ＩＬ－６组细胞数增至４．３±０．６倍，ＧＭ－ＣＳＦ组细胞数增至９．４±０．９倍；ＩＬ－６＋ＧＭ－ＣＳＦ组细胞数增至１３．７±１．０倍，明显高于对照组（Ｐ＜０．０１）。红系集落生成单位集落分析：ＩＬ－６单独应用未见ＣＦＵ－Ｅ集落形成，仅有粒－单细胞集落生成单位集落形成；ＩＬ－６＋促红细胞生成素且则ＣＦＵ－Ｅ集落数明显高于     

肾综合征出血热患者血清IL-6、尿液肿瘤坏死因子、IL-6、IL-8的检测

目的探讨细胞因子在肾综合征出血热（ＨＦＲＳ）发病中的作用。方法采用双抗体夹心ＥＬＩＳＡ法对４８例ＨＦＲＳ患者及２０例正常人血清白细胞介素（ＩＬ）－６、尿液肿瘤坏死因子（ＴＮＦ）、ＩＬ－６、ＩＬ－８进行动态检测。结果ＨＦＲＳ患者血清ＩＬ－６、尿液ＴＮＦ、ＩＬ－６、ＩＬ－８含量较对照组明显增高（Ｐ＜０．００１）；发热期已增高，低血压期继续增高，少尿期达峰值；其含量随病情加重而升高，各型间差异有显著性；血清ＩＬ－６与特异性抗体升高有明显关系，与血清β２－微球蛋白（β２－ＭＧ）、血尿素氮（ＢＵＮ）、血肌酐（Ｃｒ）均呈高度正相关；尿液ＩＬ－６与ＴＮＦ、ＩＬ－８呈显著正相关（ｒ＝０．５６２１，Ｐ＜０．００５；ｒ＝０．３８４５，Ｐ＜０．０１）。结论ＨＦＲＳ患者病程中ＴＮＦ、ＩＬ－６、ＩＬ－８均处于高活性状态，ＩＬ－６与体液免疫反应亢进所致的免疫病理损伤有关，ＩＬ－６、ＩＬ－８、ＴＮＦ参与肾脏的免疫损伤，可作为判定患者预后和转归的指标。     

健康老年人骨髓红系和粒－单系祖细胞的体外培养研究

采用造血祖细胞培养技术研究了健康老年人骨髓红系和粒－单系祖细胞的增殖能力及骨髓红系和粒－单系祖细胞分别对红细胞生成素（Ｅ_（ｐｏ））和粒－单系集落刺激因子（ＧＭ－ＣＳＦ）的反应能力。发现健康老年人红系爆式集落形成单位（ＢＦＵ－Ｅ）、红系集落形成单位（ＣＦＵ－Ｅ）和粒－单系集落形成单位（ＣＦＵ－ＧＭ）的集落数比健康非老年人明显减少；ＢＦＵ－Ｅ和ＣＦＵ－Ｅ对Ｅ_（ｐｏ）以及ＣＦＵ－ＧＭ对ＧＭ－ＣＳＦ的反应曲线低平。表明健康老年人骨髓红系和粒－单系祖细胞的增殖能力以及分别对刺激因子（Ｅ_（ｐｏ）、ＧＭ－ＣＳＦ）的反应能力减低。     

哮喘患者外周血单个核细胞中白细胞介素—6mRNA表达与气道炎症的 …

目的 探讨外周血单个核细胞（ＰＢＭＣ）中白细胞介素（ＩＬ）－６ｍＲＮＡ表达与血嗜酸细胞阳离子蛋白（ＥＣＰ）和１秒用力呼气容积占用力肺活量比值（ＦＥＶ１％）的关系。方法 对１２例过敏性哮喘发作期患者、８例缓解期患者及９例健康人，采用ＲＴ－ＰＣＲ法和图像分析半定量法检测ＰＢＭＣ中ＩＬ－６ｍＲＮＡ的表达水平及ＥＣＰ和ＦＥＶ１％。结果 发作期患者ＩＬ－６ｍＲＮＡ的表达明显高于缓解期患者和健康人（Ｐ〈０．０     

促肝细胞生长素与胰高血糖素-胰岛素治疗慢性重型肝炎疗效比较

为了比较促肝细胞生长素（ＰＨＧＦ）和胰镐血糖素－胰岛素（ＧＩ）对慢性重型肝炎的疗效，选慢重肝２６例，分Ａ、Ｂ、两组，Ａ组１４例用ＰＨＧＦ加血浆置换（ＰＥ）、自血光量子（ＵＢＩ）及综合疗法；Ｂ组１２例用ＧＩ加ＰＥ、ＵＢＩ及综合疗法。疗程均４０ｄ。结果：ＰＨＧＦ及ＧＩ疗法均能明显慢肝病死率，但两组显效率等，Ａ组优于Ｂ组。结论：ＰＨＧＦ较ＧＩ更能有效地治疗慢重 。     

In vitro effect of stem cell factor on human clonogenic marrow progenitors after myeloablative treatments

Abstract: Abnormal hematopoiesis, including a deficiency of marrow progenitors and particularly of erythroid progenitors, has been described after autologous stem cell transplantation (ASCT), persisting for several years. In order to explain this deficiency, a resistance of marrow progenitors to stem cell factor (SCF) after ASCT was investigated. Marrow samples were harvested from pregraft patients at graft collection prior to ASCT, transplanted patients 6–24 months after high-dose therapy and control patients. CD34+ cells were cultured in a serum-free clonogenic assay with increasing doses of SCF. The clonogenic efficiency without SCF was lower for BFU-E in treated groups than in controls, whereas it was not different for CFU-GM. With increasing doses of SCF a dose-dependent effect was found on the numbers of both CFU–GM and BFU–E in all groups, although the maximal number of BFU–E remained lower in treated groups. However, the SCF dose that induced 50% of maximal BFU–E growth (D₅₀) was similar in all groups. Furthermore, a dose-dependent effect on the size of BFU–E was found in all groups, with no difference in the proportion of large colonies. Thus, clonogenic erythroid progenitors from patients who have received myelotoxic treatments remain sensitive to SCF, with no evidence for a chemotherapy-related resistance.     

Insulin and insulin-like growth factor I support the proliferation of erythroid progenitor cells in bone marrow through the sharing of receptors

The effects of insulin and insulin-like growth factor I (IGF-I) on the proliferation of erythroid progenitor cells in bone marrow were studied in serum-deprived culture. Primitive human bone marrow cells were purified by cell sorting on the basis of the expression of CD34 and the Kit receptor. Insulin and IGF-I with erythropoietin (EPO) dose dependently supported the formation of erythroid colonies of CD34+/Kit+ cells in bone marrow. The direct effect of insulin and IGF-I on the stimulation of primitive erythroid progenitor cells was confirmed by single-cell proliferation studies in serum-deprived liquid suspension culture. The addition of insulin and/or IGF-I to stem cell factor (SCF) resulted in an additive increase in the number of erythroid colonies. The erythroid colonies formed by insulin and IGF-I with EPO were different in size from those formed by SCF with EPO. These findings imply that erythroid progenitor cells responding to insulin and IGF-I might be at a different developmental stage of erythropoiesis from those responding to SCF in CD34+/Kit+ cells. Similarly, insulin and IGF-I with EPO supported the proliferation of the mature erythroid progenitor cells in light-density bone marrow mononuclear cells (LDBMCs). The addition of the anti-receptor antibody to IGF-I receptor or insulin receptor partially suppressed erythroid colony formation supported with insulin or IGF-I in both CD34+/Kit+ cells and LDBMCs. The simultaneous addition of both receptor antibodies completely abrogated the erythroid colony formation. These results suggest that insulin and IGF-I directly stimulate the proliferation of the late stage of primitive erythroid progenitor cells and mature erythroid progenitor cells through the sharing of receptors.     

Clinical results of recombinant erythropoietin in transfusion-dependent patients with refractory multiple myeloma: role of cytokines and monitoring of erythropoiesis

Abstract: Recombinant erythropoietin (r-EPO) was administered to 37 patients with advanced, transfusion-dependent and chemo-resistant multiple myeloma (MM), at the fixed dose of 10,000/U s.c, 3 times a week, for 2 months. Thirteen patients (35.1%) achieved a significant response in terms of complete abolition of red cell transfusions. Factors significantly predictive of response were: a) inappropriate production of endogenous EPO, as expressed by a reduced observed/predicted ratio; b) presence of a consistant number of circulating erythroid precursors BFU–E; c) low serum levels of tumor necrosis factor (TNF) and interleukin-1 (IL-1), cytokines with inhibitory activity on erythropoiesis; d) a single line of previously received chemotherapy. Renal failure, bone marrow plasma cell infiltration, serum levels of IL-6 and other main clinical and laboratory parameters did not affect significantly the response to r-EPO. High fluorescence reticulocytes (HFR) and soluble transferrin receptor (sTfR) values were useful to detect an early stimulation of erythropoiesis in responders, while a high percentage of circulating hypochromic erythrocytes (HE), as assessed by an automated counter, identified those patients developing functional iron deficiency during r-EPO treatment. We conclude that about one-third of severely anemic patients with advanced MM, unresponsive to chemotherapy, may benefit by r-EPO therapy. The clinical management of these patients can be accomplished using non-invasive parameters, such as sTfR, HFR and HE.     

Transformation of severe aplastic anemia into acute myeloblastic leukemia with monosomy 7

 A cytogenetically normal man with severe aplastic anemia was treated with granulocyte colony-stimulating factor (G-CSF), erythropoietin (EPO), cyclosporin A, anti-thymocyte globulin, and interleukin-6 (IL-6), which resulted in a gradual improvement in his neutrophil count and hemoglobin level. After 2 years of the therapy, monosomy 7 was detected during cytogenetic analysis of his bone marrow, which evolved during a period of 5 months into acute myeloblastic leukemia. An in vitro proliferation assay of cytokine responses showed that leukemic blasts were sensitive only to G-CSF, and not to EPO or IL-6. Although allogeneic bone marrow transplantation from an HLA-matched unrelated donor was carried out in the non-remission stage, the patient died of systemic fungal infection on day 25, without any evidence of hematological engraftment. As long-term use of cytokines and immunomosuppressants in patients with severe aplastic anemia may induce or hasten the onset of a malignant transformation, careful attention must be paid to clonal evolution. Due to the poor prognosis of secondary myelodysplasia and leukemia, allogeneic bone marrow transplantation for such patients must be carried out early in the course of the disease. Received: 27 September 1995 / Accepted: 19 December 1995     

Sensitivity of erythroid progenitor colonies to erythropoietin in azidothymidine treated immunodeficient mice

The anaemia induced by 3'-azido-3'dideoxythymidine (AZT) is poorly understood. We have used a murine model of AIDS, infection of female C57BL/6 mice with LP-BM5 murine leukaemia (MuLV) virus, to determine if AZT-induced anaemia is due, in part, to decreased responsiveness of erythropoietic precursors (BFU-e) to erythropoietin (EPO). Mice in the early stage of LP-BM5 MuLV disease were given AZT in their drinking water at 1.0 and 2.5 mg/ml. AZT produced anaemia in both groups, in a dose-dependent fashion. Despite the anaemia, the number of splenic and bone marrow BFU-e in AZT treated mice increased up to five-fold over levels observed in infected untreated animals after 15 d of treatment. Colony formation by splenic and bone marrow BFUe was stimulated at lower concentrations of EPO in mice receiving AZT for 15 d than for infected, untreated mice. By day 30, sensitivity of both splenic and bone marrow BFU-e of treated animals returned to that observed from cells of infected untreated animals. The mean plasma levels of EPO observed in AZT treated mice were appropriate for the degree of anaemia observed when compared with phenylhydrazine (PHZ) treated mice. The numbers of BFU-e and the percentage of bone marrow erythroblasts observed were comparable in AZT and PHZ treated mice with similar degrees of anaemia. However, reticulocytosis was inappropriate for the degree of anaemia observed in AZT treated infected mice. AZT-induced peripheral anaemia in the face of increased numbers of BFU-e and increased levels of plasma EPO suggest a lesion in terminal differentiation.     

肝硬化患者骨髓促红细胞生成素水平的检测

本文采用放射免疫方法检测肝硬化患者的血清和骨髓中的红细胞生成素（EPO）含量。研究结果显示,患者血清中EPO水平与健康人相比,显著升高（P＜0．001）,普遍存在EPO应答障碍,致使贫持续存在。与此相反,患者骨髓EPO水平低于正常健康人（P＜0．001）。提示,肝硬化患者存在骨髓造血机能障碍。     

Reversible suppression of bone marrow response to erythropoietin in Plasmodium falciparum malaria

To study the importance of bone marrow inhibition in the pathogenesis of malarial anaemia, haematological and parasitological parameters were followed in patients with acute malaria. Three patient categories were studied, severe malarial anaemia (SA), cerebral malaria (CM) and uncomplicated malaria (UM). Red cell distribution width (RDW) was used as a surrogate marker of release of young erythrocytes and reticulocytes. Initially RDW was low in all patients in spite of markedly increased concentrations of erythropoietin (EPO). 3 d after institution of treatment and coinciding with parasite clearance RDW increased dramatically, reaching the highest levels 1–2 weeks later. Although severe anaemia was corrected by blood transfusion during the first 3 d of treatment, the peak RDW correlated significantly with the initial EPO levels. This suggests that Plasmodium falciparum infection causes a rapidly reversible suppression of the bone marrow response to EPO. Furthermore, the inhibition of bone marrow response was a general finding irrespective of initial haemoglobin levels suggesting that the severity of anaemia depends upon the degree of peripheral erythrocyte destruction in patients with suppressed bone marrow response to EPO.     

Erythropoietin improves cardiac function through endothelial progenitor cell and vascular endothelial growth factor mediated neovascularization.

AIMS: Erythropoietin (EPO) improves cardiac function and induces neovascularization in chronic heart failure (CHF), although the exact mechanism has not been elucidated. We studied the effects of EPO on homing and incorporation of endothelial progenitor cells (EPC) into the myocardial microvasculature and myocardial expression of angiogenic factors. METHODS AND RESULTS: CHF was induced in rats by coronary artery ligation resulting in myocardial infarction (MI) after bone marrow had been replaced by human placental alkaline phosphatase (hPAP) transgenic cells. We studied the effects of darbepoetin alfa treatment (EPO, 40 microg/kg, every 3 weeks, starting 3 weeks after MI) on longitudinal changes in left ventricular (LV) function, circulating EPC, myocardial histology, and expression of vascular endothelial growth factor (VEGF) determined 9 weeks after MI. EPO prevented LV-dilatation and improved cardiac function (all P < 0.05), which was associated with 42% increased capillary growth (P < 0.01). EPO-induced mobilization of EPC from the bone marrow (P < 0.01), which resulted in a three-fold increased homing of EPC into the cardiac microvasculature. The percentage of the endothelium that consisted of bone marrow derived cells was significantly increased (3.9 +/- 0.5 vs. 11.4 +/- 1%, P < 0.001) comprising 30% of the newly formed capillaries. In addition, EPO treatment resulted in a 4.5-fold increased myocardial expression of VEGF, which correlated strongly with neovascularization (r = 0.67; P < 0.001). VEGF was equally expressed by endothelial cells of myocardial and bone marrow origin. CONCLUSION: EPO-induced neovascularization in post-MI heart failure is mediated through a combination of EPC recruitment from the bone marrow and increased myocardial expression of VEGF.     

Increase of CD34 positive cells in polycythaemia vera

Abstract: The purpose of the present work was to evaluate the proliferative character of polycythaemia vera (PV). Therefore, in 15 patients with different stages of PV we assessed the level of CD34 positive (CD34+) cells in peripheral blood and bone marrow, erythroid colony growth of bone marrow cells and plasma erythropoietin (EPO). The mean concentration of CD34+ cells in blood was significantly increased in PV patients (9.0±11.2×10³/mL) compared to healthy controls (2.0±1.7×10³/mL). In aspirated bone marrow no such difference between PV and control subjects was present. Six patients with splenomegaly and/or requirement for chemotherapy had significantly higher mean blood levels of CD34+ cells compared to the remaining PV patients. All PV patients presented EPO independent erythroid colonies. Three PV patients with anaemia and long disease duration had high EPO levels.     

Stimulation of human BFU(E) by products of human monocytes and lymphocytes

Regulation of normal human peripheral blood BFU(E) by lymphocytes and monocytes was investigated using plasma clot cultures. Monocyte depletion from peripheral blood mononuclear cells (MNC) resulted in 28 to 98% inhibition of BFU(E) growth in 16 of 17 experiments, but T lymphocyte depletion caused no decrease in BFU(E). When both T cells and monocytes were removed there was an 80% decrease in BFU(E) growth. Addition of as few as 10(4) monocytes alone increased BFU(E) to 88% of expected levels, but addition of 5 x 10(4) T lymphocytes resulted in an increase to only 57% of anticipated BFU(E) numbers. Conditioned media (CM) from 5 x 10(5) to 10(6) unstimulated T lymphocytes per ml stimulated BFU(E) growth from cultures of non-adherent cells, but not from cultures of T cell depleted MNC. Monocyte CM produced peak (2.2-fold) stimulation with only 10(5) monocytes/ml. CM from 5 x 10(5) to 10(6) monocytes/ml had no detectable erythroid burst promoting activity (BPA). Thus, unstimulated monocytes produce BPA, and at high cell concentrations, also produce a presumed inhibitor of BPA production or biologic activity. Although T cells may play some role in control of BFU(E), monocytes appear to be critical for optimal BFU(E) growth in vitro.