阵发性睡眠性血红蛋白尿症患者骨髓正常细胞及异常克隆分析

目的 分析阵发性睡眠性血红蛋白尿症（PNH）患者骨髓正常细胞（CD59^ )及异常克隆细胞（CD59^－）的细胞构成。方法 骨髓有核细胞免疫荧光标记后用流式细胞术分析。结果 （1）PNH患者骨髓正常及异常克隆的细胞组成在有核细胞水平是显著不均衡的，正常的CD59^ 细胞以淋巴细胞窗内细胞为主，异常的CD59^－细胞则以粒细胞窗及原始细胞窗内细胞为主。（2）PNH患者骨髓CD59^ 正常细胞群所含CD59^ 细胞总数较异常者显著减少，淋巴细胞窗及粒细胞窗内CD59^ 细胞均较异常者显著减少，前者减少更为显著。结论 PNH患者骨髓正常细胞及异常克隆的细胞构成在有核细胞水平及CD59^ 细胞水平都有显著差异，正常造血呈衰竭状态。因此，在考虑用患者自体骨髓体外净化去除异常克隆后回输给患者重建造血时，应对正常造血细胞作进一步处理，否则正常造血细胞可能不具备重建造血的能力。     

阵发性睡眠性血红蛋白尿症CD34+CD59+细胞长期造血的动物实验

目的获得阵发性睡眠性血红蛋白尿症（PNH）患者正常克隆干细胞（CD34^＋CD59^＋）支持长期造血的证据。方法纯化、扩增PNH患者骨髓CD34^＋CD59^＋细胞并移植入联合免疫缺陷（SCm）小鼠体内,90d后以首次移植的小鼠骨髓细胞进行二次移植。结果分别移植了PNH患者和正常对照骨髓CD34^＋CD59^＋细胞的小鼠在移植后第90天,外周血细胞水平均恢复正常;两组小鼠的组织中均有CD45的表达,且差异无统计学意义。分别移植了由PNH患者和正常对照骨髓CD34^＋CD59^＋细胞重建造血的小鼠骨髓细胞的小鼠在移植后第30天,外周血细胞水平差异无统计学意义;两组小鼠的组织中均有C1M5的表达,且差异无统计学意义;移植后小鼠骨髓细胞里均可扩增出供者的SRY基因片段。结论经体外纯化、扩增后的PNH患者骨髓CD34^＋CD59^＋细胞具有与正常CD34^＋CD59^＋细胞同样的长期造血的能力。     

阵发性睡眠性血红蛋白尿症临床分析

本文报道15例阵发性睡眠性血红蛋白尿症的临床表现，骨髓、溶血试验和并发症等，并阐述阵发性睡眠洼血缸蛋白尿症的治疗、转归和发作诱因等。     

阵发性睡眠性血红蛋白尿症患者外周血CD55、CD59测定

阵发性睡眠性血红蛋白尿症(paroxysmal nocturnal hemoglobinuria,PNH)是一种获得性克隆性造血干细胞疾病,临床表现为全血细胞减少、血管内溶血、反复静脉血栓形成[1],PNH是由于各种原因造成骨髓损伤,导致糖化肌醇磷脂-锚(PIG-A)基因突变,使糖基磷酯酰肌醇(GPI)锚连蛋白合成障碍,患者血细胞呈现多种锚连接蛋白的减少或缺失,其中包括衰变加速因子(DAF,CD55)和反应性溶血膜抑制物(MIRL,CD59)的缺失[2].作者测定了PNH患者外周血CD55、CD59的表达,以探讨CD55、CD59检测对PNH诊断及鉴别诊断的意义.     

CD55、CD59在阵发性睡眠性血红蛋白尿诊断中的作用

目的：探讨衰变加速因子(CD55)、反应性溶血膜抑制物(CD59)在阵发性睡眠性血红蛋白尿(PNH)诊断中的意义。方法：用流式细胞仪检测CD55、CD59的表达量(％)。结果：PNH组CD55、CD59明显低于AA—PNH组和AA组、对照组。结论：检测患者外周血粒细胞膜表面抗原CD55、CD59可作为PNH的早期确诊指标,其中CD59是较敏感指标,CD55是特异性指标。     

牡丹江地区10年阵发性睡眠性血红蛋白尿症流行病学调查分析

自1985年1月1日至1994年末在牡丹江地区4市（县）、共调查了16068693人次，发现PNH33例，标化发病率为2.85／106，明显高于前10年的调查，标化患病率为9.1／106，明显高于发病率。边境发病率较高，人口密集、工业集中的城市发病率低。多发于中青年，男多于女。睡眠、感染、过劳、服用某种药物易诱发本病。     

阵发性睡眠性血红蛋白尿症对症治疗的评价

目的：评价对PNH的对症治疗。方法：分析本院274例PNH的随访转归结果与对症治疗的关系。结果：能够坚持较长时期对症治疗者比不能坚持治疗者的生存期延长。结论：能够坚持较长时期对症治疗者可获得较长生存期及从事某些工作的能力，其中10．6％可获得自发性临床缓解。     

阵发性睡眠性血红蛋白尿患者骨髓多向祖细胞的特点

采用造血祖细胞体外培养技术,研究阵发性睡眠性血蛋白尿（ＰＮＨ）患者骨髓多向祖细胞（ＣＦＵ－Ｍｉｘ）的增殖能力;骨髓细胞经酸化ＡＢ血清处理后ＣＦＵ－Ｍｉｘ的增殖能力。发现ＰＮＨ患者骨髓ＣＦＵ－Ｍｉｘ的集落数明显低于正常;骨髓细胞顷新鲜酸化ＡＢ血清处理后的ＣＦＵ－Ｍｉｘ集落数,明显低于经热酸化ＡＢ血清处理后培养的集落数。     

CD16在阵发性睡眠性血红蛋白尿症中的特异性表达

目的:检测中性粒细胞表面分化抗原CD16在阵发性睡眠性血红蛋白尿症(PNH)中表达,评价其在PNH诊断中的意义。方法:运用流式细胞术检测40名健康体检者及36例初治PNH患者外周血中性粒细胞表面分化抗原CD16,并与CD55、CD59及与传统Ham's试验结果进行比较研究。结果:PNH患者外周血细胞的CD16、CD55及CD59均有不同程度的缺乏,与健康对照组比较,差异有统计学意义(P<0.01);与传统Ham's试验比较其灵敏度高,特异度高。结论:细胞膜上糖基化磷脂酰肌醇锚的锚定蛋白CD16的缺陷与PNH有密切关系,流式细胞术检测PNH患者外周血中性粒细胞CD16可作为诊断PNH的有效辅助检查方法,具有一定临床意义。     

阵发性睡眠性血红蛋白尿症患者大颗粒淋巴细胞变化的研究

检测了３２例正常人，３２例阵发性睡眠性血红蛋白尿症（ＰＮＨ）患者外周血大颗粒淋巴细胞（ＬＧＬ）计数，发现：①ＰＮＨ患者外周血ＬＧＬ百分率及绝对值均显著低于正常对照组（Ｐ＜０．００１）。②发作期ＰＮＨ患者外周血ＬＧＬ百分率及绝对值均显著低于缓解期（Ｐ＜０．００１）。③缓解期ＰＮＨ患者外周血ＬＧＬ百分率及绝对值与正常对照组比较均无显著性差异（Ｐ＞０．５）。结果提示：可以把ＬＧＬ计数的变化作为判断ＰＮＨ患者预后及疗效的指标。     

Association analysis of cytokine polymorphisms and plasma level in Northern Chinese Han patients with paroxysmal nocturnal hemoglobinuria

Background  While the incidence of paroxysmal nocturnal hemoglobinuria (PNH) is relatively high in Northern China, the exact mechanism of the disease remains unknown. Immunoregulatory cytokine polymorphisms can directly regulate the expression levels of cytokines, which play a crucial role in many diseases. The purpose of this study was to study cytokine gene single nucleotide polymorphisms (SNPs) and the correlated cytokine expression levels in relationship to the PNH pathogenesis.

Methods  Peripheral blood samples were collected from 30 PNH patients and 40 healthy donors; all of the samples were collected from the Han people of Northern China. Eight SNP loci in five cytokine genes, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β), interleukin-6 (IL-6), and IL-10, and aplastic anemia (AA) were assessed. TNF-a, TGF-b, IFN-g, IL-6, and IL-10 were analyzed by sequence-specific primer polymerase chain reaction (PCR-SSP). The plasma protein levels of TNF-a, TGF-b, and IFN-g were assessed by an ELISA.

Results  The PNH patients had a lower frequency of the TC/GG genotype of the TGF-b gene (P <0.01) and a higher frequency of the C allele in the TGF-b gene (+10) compared to the controls (P <0.05). The predominant genotype of the +874 locus of the IFN-g gene was TA in the PNH patients, while that in the predominant genotype was AA in the control group and was statistically significant (P <0.001). The frequency of the T allele in the IFN-g gene was dramatically higher in the PNH patients than in the controls (P <0.05). The PNH patients had a reduced frequency of the GC and CC genotypes, as well as the C allele at locus –174 of the IL-6 gene compared to the controls (P <0.01). In addition, the plasma concentrations of TNF-a, TGF-b, and IFN-g were significantly higher in the PNH group compared to the control group (P <0.01).

Conclusions  Expression levels of the TNF-a, TGF-b, and IFN-g cytokines play an important role in PNH. The GC and CC genotypes, as well as the C allele of the IL-6 gene may protect the Han people of Northern China against PNH. Additionally, the TC/GG genotype of the TGF-b gene may be the protective allele. In contrast, the TA genotype and the T allele for the IFN-g gene, as well as the C allele of TGF-b may be susceptible to PNH. However, SNPs in the TNF-a and IL-10 genes did not correlate with PNH development. Alternatively, the increased plasma concentrations of TNF-a, TGF-b, and IFN-g in PNH patients may also be related to PNH development.

     

Successful application of nonmyeloablative stem cell transplantation for paroxysmal nocturnal hemoglobinuria

Paroxysmal nocturnal hemoglobinuria （PNH） is a consequence of nonmalignant clonal expansion of one or several hematopoietic stem cells that have acquired a somatic mutation of phosphatedalinositol glycae-class A （PIGA） gene leading to deficient glycosyl phosphatidylinositol （GPI） synthesis. The pathophysiology of PNH is an abnormality in the glycolipid-anchored proteins of the membranes of blood cells. The absence of complement control proteins, including decay accelerating factor （DAF, CD55） and membrane inhibitor of reactive lysis （MIRL, CD59）, results in intravascular lysis of the abnormal cells. Patients with PNH manifest not only clinical and laboratory signs of chronic hemolytic anemia, but also formation of cytopenia and venous thromboses.[第一段]     

Successful management of unstable angina in a ravulizumab-treated patient with paroxysmal nocturnal hemoglobinuria

Ravulizumab is an anti-C5 antibody approved for treating paroxysmal nocturnal hemoglobinuria (PNH). In August 2019, a 77-year-old Japanese man with PNH, who had been on ravulizumab treatment for 2 years, was hospitalized for chest discomfort and malaise. Electrocardiography identified a right bundle block, and elevated serum troponin I and d-dimer suggested ischemic heart disease. Cardiac catheterization revealed severe stenosis in the left anterior descending coronary artery, and intracoronary stenting relieved his chest discomfort. The final diagnosis was unstable angina unrelated to ravulizumab, and the patient’s ravulizumab treatment was uninterrupted with no significant complications of PNH. This case report highlights the importance of continuing complement inhibition therapy during acute coronary events.     

近十年78例新发阵发性睡眠性血红蛋白尿症的临床分析

目的　研究九十年代阵发性睡眠性血红蛋白尿症（PNH）患者的临床特征,以增加对PNH新的认识,指导PNH的诊断及治疗.方法　对1990年1月至1999年11月我院诊断的78例新发PNH患者的临床表现、实验室检查及治疗情况进行了回顾性分析.结果　①与八十年代的病例相比,新发PNH患者的发病年龄有所增大（中位年龄由27岁升至34岁）；女性病例增多（由18.5%升至38.5%）；无血红蛋白尿发作病例增多（由24.2%升至38.5%）；血栓形成发生率仍较低（3.0%对6.4%）. ②所有患者均无家族遗传史. ③部分患者表现类似骨髓增生异常综合征：骨髓有2-3系病态造血（19.2%）,染色体核型异常（12.2%）,姊妹染色单体分染阴性（8.9%）；部分患者表现类似再生障碍性贫血：骨髓增生减低（12.3%）,Ham's试验阴性（34.2%）.但100.0%（25/25例）患者外周血红细胞及粒细胞CD55或CD59表达异常,可作为鉴别诊断的特异性指标.④83.8%患者对肾上腺糖皮质激素为主的治疗有效,但易复发,1年复发率为54.2%.8例对肾上腺糖皮质激素治疗效果差或有依赖的难治性、复发性PNH用低剂量化疗（MP方案：马法兰2-6?mg*d-1；强的松0.5?mg*kg-1*d-1）治疗有效率为62.5%,而且均未出现严重的骨髓抑制及其它毒副反应.结论　PNH作为一种后天获得性疾病,更多见于成年男性.CD55、CD59的检测有助于提高PNH的检出率,大部分的PNH患者对肾上腺糖皮质激素为主的治疗反应良好,对难治性、复发性PNH患者可试用低剂量化疗.     

近十年78例新发阵发性睡眠性血红蛋白尿症的临床分析

目的　研究九十年代阵发性睡眠性血红蛋白尿症 (PNH)患者的临床特征 ,以增加对PNH新的认识 ,指导PNH的诊断及治疗。方法　对 1990年 1月至 1999年 11月我院诊断的 78例新发PNH患者的临床表现、实验室检查及治疗情况进行了回顾性分析。结果　①与八十年代的病例相比 ,新发PNH患者的发病年龄有所增大 (中位年龄由 2 7岁升至 34岁 ) ;女性病例增多 (由 18 5 %升至 38 5 % ) ;无血红蛋白尿发作病例增多 (由 2 4 2 %升至 38 5 % ) ;血栓形成发生率仍较低 (3 0 %对 6 4 % )。②所有患者均无家族遗传史。③部分患者表现类似骨髓增生异常综合征 :骨髓有 2 - 3系病态造血(19 2 % ) ,染色体核型异常 (12 2 % ) ,姊妹染色单体分染阴性 (8 9% ) ;部分患者表现类似再生障碍性贫血 :骨髓增生减低 (12 3% ) ,Ham’s试验阴性 (34 2 % )。但 10 0 0 % (2 5 / 2 5例 )患者外周血红细胞及粒细胞CD5 5或CD5 9表达异常 ,可作为鉴别诊断的特异性指标。④ 83 8%患者对肾上腺糖皮质激素为主的治疗有效 ,但易复发 ,1年复发率为 5 4 2 %。 8例对肾上腺糖皮质激素治疗效果差或有依赖的难治性、复发性PNH用低剂量化疗 (MP方案 :马法兰 2 - 6mg·d 1;强的松 0 5mg·kg 1·d 1)治疗有效率为 6 2 5 % ,而且均未出现严重的骨髓抑     

Clinical analysis of 78 cases of paroxysmal nocturnal hemoglobinuria diagnosed in the past ten years

Objective To learn more about the clinical and laboratory features of patients with paroxy smal nocturnal hemoglobinuria (PNH) diagnosed in the past ten years. Methods Clinical and laboratory data for 78 cases of PNH diagnosed from January 1990 to November 1999 in our hospital were analyzed retrospectively. Results In comparison with PNH cases reported in the 1980s, the newly diagnosed PNH cas es showed the following features: (1) older age of disease onset (from 27 to 34 years); more female cases (from 18. 5% to 38. 5%); more cases without hemoglobin uria (from 24. 2% to 38. 5%). (2) No positive family hereditary history. (3) Bone marrow dysplasia, abnormal karyotype and negative sister chromatid differen tiation were found in 19. 2%, 12. 2% and 8. 9% of the PNH patients, respectively . 12. 3% of the patients had bone marrow hypoplasia, and most of them had no he moglobinuria. Ham’s tests were negative in about 34. 2% of the cases. CD55 and CD59 on peripheral blood cells were deficient in 100. 0% of the cases, suggesti ng that CD55 and CD59 tests can improve the diagnosis of PNH. (4) Adrenocortic al hormone was effective in 83. 8% of the patients, 54. 2% of whom relapsed with in one year. Eight refractory and relapsed patients were treated with low dose chemotherapy (MP therapy: Melphalan 2-6 mg·d; Prednisone 0. 5 mg·kg(-1) ·d(-1) ). Five (62. 5%) of them showed positive responses. Bone marrow failure and other side effects were not serious in this group of patients Conclusions PNH, an acquired blood disease seen more often among adult males, can be diagno sed more sensitively by hemocyte member CD55 and CD59 tests and treated more eff ectively with adrenocortical hormone or low dose chemotherapy.     

Inhibiting effects of low- molecular weight heparin and adrenocortical hormone on hemolysis of red cells in patients with paroxysmal nocturnal hemoglobinuria in vitro

Objective To study the effects of low- molecular weight heparin (LMWH) and adrenocortical hormone (dexamethasone) on the hemolysis of red cells of patients with paroxysmal nocturnal hemoglobinuria (PNH) in vitro.Methods Using Ham’s test and micro- complement lysis sensitive test (mCLST), the changes in hemolysis of red cells from 6 typical PNH cases were examined after adding LMWH and dexamethasone in different concentrations into the test solution in vitro. The effects of LMWH and dexamethasone on the coagulation of the tested blood samples were also studied using the activated partial thromboplastin time (APTT) test. Results Both LMWH and dexamethasone inhibited the hemolysis of PNH red cells, and they also showed a synergistic effect. The inhibiting effects were dose- dependent. Moreover, a tolerable dose of LMWH induced a limited prolongation of APTT. Dexamethasone showed two possible mechanisms in the inhibition of PNH red cells hemolysis through Ham’s test and mCLST, respectively: ① inhibiting both antibodies binding to red cells and ② the initiation of the activation of complement 3 (C3). LMWH could inhibit hemolysis as determined by both Ham’s test and mCLST, which indicated that LMWH could block the activation of complement cascade. Conclusions Both LMWH and dexamethasone could inhibit hemolysis in PNH, and they showed a synergistic effect. Their mechanisms of inhibiting hemolysis differed from each other. Furthermore, a tolerable dose of LMWH induced a limited prolongation of APTT. LMWH might be useful for controlling acute hemolysis in patients with PNH and reducing the dose of adrenocortical hormone.